CN105886428B - One plant of Streptomycesalbidoflhaving and its application in microbial manure - Google Patents
One plant of Streptomycesalbidoflhaving and its application in microbial manure Download PDFInfo
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- CN105886428B CN105886428B CN201610207349.1A CN201610207349A CN105886428B CN 105886428 B CN105886428 B CN 105886428B CN 201610207349 A CN201610207349 A CN 201610207349A CN 105886428 B CN105886428 B CN 105886428B
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/465—Streptomyces
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
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- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F3/00—Fertilisers from human or animal excrements, e.g. manure
Abstract
The invention discloses one plant to have the Streptomycesalbidoflhaving W68 (bacterial strain deposit number is CGMCC No.12210) for being isolated from deep-sea sludge of broad-spectrum antifungal activity and using the microbial bacterial agent of bacterium preparation and using the bacterium as the application technology of the microbial manure of material.Microbial bacterial agent and be not only simple using the bacterium as the application technology of the microbial manure of material, efficiently prepared by the Streptomycesalbidoflhaving W68 provided by the invention to be isolated from deep-sea sludge, and it is environmentally friendly, and the bacteria preparation spore concentration of solid fermentation can achieve 10,000,000,000.It is confirmed through pot experiment, the Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge shows apparent control effect to the prevention and treatment of the silborne fungal diseases such as root rotof flax, Muskmelon Fusarium wilt, phytoph-thora capsici leonian and take-all.The present invention has huge application prospect for the green prevention and control of silborne fungal diseases.
Description
Technical field
The present invention relates to micro- Bai Huanglian of one plant of source Yu Haiyang with broad-spectrum antifungal activity and biological and ecological methods to prevent plant disease, pests, and erosion application potential
Mould (Streptomyces albidoflavus) bacterial strain W68, belongs to application and crop disease control of marine microorganism etc.
Microorganisms technical field.
Background technique
Fungal disease is the main reason for causing crop yield and quality to reduce.The frequent use of chemical pesticide is led
It causes some disease fungus drug resistances sharply to enhance, cause the pollution of environment and agricultural product, and destroys microorganism inside and outside plant
Balance and aggravate the eruption and prevalence etc. of disease.
The study on prevention that soil passes fungal diseases of plants is a hot and difficult issue all the time, and fungicide on the market is general
For to its without or have little effect.People have also screened the biocontrol microorganisms of some anti-soil fungis for many years, as shield shell is mould
(Coniothyrium minitans) is used for sheath blight fungus and Pythium ultimum for the prevention and treatment of sheath blight fungus, Trichoderma viride
The prevention and treatment of seedling diseases caused by (Pythium ultimum) or the various diseases including root rot, streptomyces griseoviridus
(Streptomyces griseoviridis) is used for the prevention and treatment etc. of cotton wilt.Though these soil fungi disease biocontrol bacterial strains
So all there is stronger antifungal activity, but field test effect is poor, many fermenting performances are poor, and in the application
Usually there is certain region limitation.
Summary of the invention
The purpose of the present invention is to provide one plant to have the active Streptomycesalbidoflhaving of broad-spectrum disease resistance fungal pathogens
(Streptomyces albidoflavus) bacterial strain W68 and its application in prevention and treatment silborne fungal diseases.
In fact, the present invention relates to one plant of Streptomycesalbidoflhaving (Streptomyces albidoflavus) bacterial strain W68.
It is related to the microbial inoculum that active constituent is Streptomycesalbidoflhaving (Streptomyces albidoflavus) bacterial strain W68.
It is related to Streptomycesalbidoflhaving (Streptomyces albidoflavus) bacterial strain for preventing and treating soil-borne fungus disease
The microbial inoculum of W68.
It is related to Streptomycesalbidoflhaving (Streptomyces albidoflavus) bacterial strain W68 as microbial manure anti-
Control the application in silborne fungal diseases.
Protected be isolated from deep-sea sludge Streptomycesalbidoflhaving W68 and with the bacterium preparation microorganism formulation and
Using the bacterium as the application technology of the microbial manure of material.
Streptomycesalbidoflhaving W68 in deep-sea of the present invention is located away from marine active sludge, and quantitative activated sludge is filled
Enter and pre-processed in the sterile water containing small bead, using the Gause I for containing final concentration of 50 μ g/ml potassium bichromate
Culture medium is separately cultured, and is reused PDA culture medium and is carried out purifying culture.16S rDNA sequence is compared in ncbi database
Column, identify the Pseudomonas in Streptomyces albidoflavus monoid.The bacterium is preserved on March 14th, 2016 at present
China Committee for Culture Collection of Microorganisms's common micro-organisms center, address are as follows: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Number (CGMCC), deposit number are CGMCC No.12210
The biological property of the heretofore described Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge is as follows:
The colony characteristics of 1. Streptomycesalbidoflhaving W68 of table
| Culture medium | The positive face color of bacterium colony | Bacterium colony back side color |
| ISP2 | White | Yellow |
| ISP3 | Micro- Huang | Micro- Huang |
| ISP4 | White | White |
| ISP5 | Microscratch growth | Non-pigment |
| Bennete | White | Micro- Huang, white |
| Cazpek’s | Micro- Huang | Micro- Huang |
| Glu+Asp | Microscratch growth | Non-pigment |
| Glu+Asp+M | White | White |
| PDA | White | Yellow |
Colony characteristics of the Streptomycesalbidoflhaving W68 on various culture mediums are shown in Table 1.Average incubation time is 5-7 days.?
In PDA culture medium, aerial hyphae initial stage is white, powdery, there is white and the secondary clump of creamy.Micro- Huang is presented in bacterium colony after producing spore
Color.Scanning electron microscopic observation fibrillae of spores contains about 30 to 50 spores at flexure type, no spiral, every fibrillae of spores；Spore is in cylinder
Shape, surface is smooth, no protrusion, no thorn (Fig. 1).
The antifungal activity of the heretofore described Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge is as follows:
The Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge has the antifungal activity of wide spectrum.Plate face-off experiment display,
Streptomycesalbidoflhaving W68 is to fusarium verticillioides (Fusarium verticillioides), Fusarium oxysporum (Fusarium
Oxysporum), Bipolaris sacchari (Bipolaris sorokiniana), Rhizoctonia solani Kuhn (Rhizoctonia solani),
Verticillium dahliae bacterium (Verticillium dahliae), grey big angle base shell bacterium (Magnaporthe grisea) etc. cause
A variety of disease fungus of plant disease all have good antagonism (Fig. 2).
The biological and ecological methods to prevent plant disease, pests, and erosion application technology and method of the heretofore described Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge is as follows:
(1) seed coating technique is used for the microorganism live bacteria preparation of preparation；
(2) using Streptomycesalbidoflhaving W68 as the microbial bacterial agent of material or microbial manure；
(3) using Streptomycesalbidoflhaving W68 as material, using common agricultural wastes as solid fermentation material, spore can be obtained
Sub- concentration is 1 × 1010The microbial bacterial agent or microbial manure of a/g；
(4) microbial bacterial agent or microbial manure purposes can be used for being coated, soak seed, root dipping, cave are applied, ditch spread, base manure use.
The biological and ecological methods to prevent plant disease, pests, and erosion application effect of the heretofore described Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge is as follows:
The Streptomycesalbidoflhaving W68 of deep-sea sludge is isolated to root rotof flax (Bipolaris sorokiniana)
Control effect.In the pot experiment prevented and treated root rotof flax, using the coated method of seed, Streptomycesalbidoflhaving is used
W68 bacteria preparation improves emergence rate of the wheat seed in the soil containing Cochliobolus sativus, and plant height, plant weights are above
The control of morbidity group, and can achieve level of growth (Fig. 3) of the wheat seed in no pathogen soil substantially.
It is isolated from prevention and control of the Streptomycesalbidoflhaving W68 of deep-sea sludge to Muskmelon Fusarium wilt (Fusarium oxysporum)
Effect.By seed coating and root irrigation, can significantly be overcome by fusarium wilt using Streptomycesalbidoflhaving W68 bacteria preparation
Caused continuous cropping obstacles: muskmelon is continuously planted in the field that last year kind crosses muskmelon, the muskmelon shoot survival percent of processing group can
To reach 93%, the shoot survival percent of blank control group is only 13% (Fig. 4).
The Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge prevents phytoph-thora capsici leonian (Phytophthora capsici)
Control effect.Seedling is handled using Streptomycesalbidoflhaving W68 bacteria preparation root dipping when pepper seedling is transplanted, can significantly mitigate Phytophthora capsici
The occurring degree of disease.Compared with the control group, the processed plant strain growth cycle stretch-out of root dipping about 1-2 weeks, chilli yield increase
5% (Fig. 5).
The Streptomycesalbidoflhaving W68 of deep-sea sludge is isolated to take-all (Gaeumannomyces.graminis)
Control effect.The use of deep-sea Streptomycesalbidoflhaving W68 active bacteria formulation can significantly reduce the diseased plant rate of wheat heading stage, drop
Low disease index.Compared with chemical agent, the preventive effect of Streptomycesalbidoflhaving W68 active bacteria formulation is substantially better than chemical reagent triazole
Ketone is suitable with the net preventive effect of total eclipse.It can be seen that Streptomycesalbidoflhaving W68 active bacteria formulation can substitute the use of chemical agent completely
(Fig. 6).
Detailed description of the invention:
Fig. 1 is growthform and scanning electron microscopic observation figure of the Streptomycesalbidoflhaving W68 in PDA culture medium.
Fig. 2 is antagonism figure of the Streptomycesalbidoflhaving W68 to various plants pathogen.
Fig. 3 is the results from pot experiment test figure of Streptomycesalbidoflhaving W68 prevention and control root rotof flax.
Fig. 4 is the field experiment result figure of Streptomycesalbidoflhaving W68 prevention and control Muskmelon Fusarium wilt.
Fig. 5 is the field experiment result figure of Streptomycesalbidoflhaving W68 prevention and control phytoph-thora capsici leonian.
Fig. 6 is the field experiment result figure of Streptomycesalbidoflhaving W68 prevention and control take-all.
Specific embodiment
Embodiment below facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method is unless otherwise specified conventional method.Test material as used in the following examples is unless otherwise specified certainly
What routine biochemistry reagent shop was commercially available.
In following implementing regulations, following culture medium is used:
Gause I screening and culturing medium: soluble starch 20g, KNO31g, K2HPO40.5g, MgSO4·7H2O 0.5g,
NaCl 0.5g, FeSO4·7H2O 0.01g, NaCl 0.5g, agar 20g, tap water constant volume to 1000mL, pH 7.2~7.4.
PDA culture medium: potato dextrose agar, potato 200g filter to take filtrate, Portugal after boiling 30 minutes
Grape sugar 20g, agar 15-20g, tap water constant volume to 1000ml, natural pH；115 DEG C of high-temperature sterilizations.
ISP2 culture medium (yeast extract malt extract agar): yeast extract 10g, glucose 4g, malt extract 10g, agar 20g, from
Water constant volume is to 1000mL, pH 7.3.
ISP3 culture medium (oatmeal agar): oatmeal 20g immersion liquid, mark amount salting liquid 1ml, agar 20g, tap water constant volume
To 1000mL, pH 7.2.
ISP4 culture medium (starch agar): soluble starch 10g, MgCO31g, K2HPO41g, NaCl 1g, NaNO31g,
Agar 15g, tap water constant volume to 1000ml, pH 7.2~7.4.
ISP5 culture medium (glycerol asparagine agar): L- asparagine 1g, K2HPO41g, mark amount salting liquid 1ml, glycerol
10g, agar 20g, tap water constant volume to 1000ml, pH 7.0~7.4
Bennete culture medium: glucose 10g, beef extract 1g, agar 15g, yeast extract 1g, hydrolyzed casein 2g, tap water are fixed
Hold 1000mL, pH 7.3
Cazpek ' s culture medium (Cha Shi): sucrose 30g, NaNO3 2g,K2HPO4 1g,MgSO4·7H2O 0.5g, KCl
0.5g, FeSO4·7H2O 0.01g, agar 20g, tap water constant volume to 1000mL, pH 7.2~7.4.
Glu+Asp culture medium (glucose asparagine agar): glucose 10g, asparagine 0.5g, K2HPO40.5g,
Agar 15g, tap water constant volume to 1000mL, pH 7.2~7.4.
Glu+Asp+M culture medium (glucose asparagine meat extract agar): glucose 10g, asparagine 0.5g, beef extract
2g, K2HPO40.5g, agar 15g, tap water constant volume to 1000mL, pH 6.8.
Mark amount salting liquid: FeSO4·7H2O 0.1g, ZnSO4·7H2O 0.1g, MnCl2·4H2O 0.1g, distilled water are fixed
Hold to 100ml.
2 × YT fluid nutrient medium: peptone 16g, yeast powder 10g, NaCl 5g, tap water constant volume to 1000mL are natural
pH。
Embodiment 1
Separation, identification and the biological characteristics of bacterial strain
One, the separation and purifying of bacterial strain
This experiment isolated several plants of actinomyces, process from marine active sludge is as follows: 1g sample is added on sterilizing
The 300ml triangular flask equipped with small bead and 99ml sterile saline in, 28 DEG C of shaking table culture 1h.L ml suspension is taken, is adopted
10 are diluted to the method for gradient dilution-4, take stoste, 10-2Dilution and 10-4Dilution is respectively coated potassium bichromate final concentration
For 50 μ g/ml Gause I culture mediums, respectively at the 5th day, the 7th day and the 10th day picking single colonie, then plate streak is used
In being purified and be separately cultured in PDA culture medium.
Gained purifying bacterial strain is saved in -80 DEG C using 20% glycerol.
Two, the identification of bacterial strain
1, Morphological Identification
It is isolated from the Morphological Identification of the Streptomycesalbidoflhaving W68 of deep-sea sludge, with the method for plate streaking in different trainings
Morphologic observation (table 1) is carried out after supporting base inoculated and cultured.
2, Molecular Identification
PCR amplification 16S rDNA uses universal primer 27F and 1492R, and (annealing temperature is about for 56 DEG C, target sequence
1400bp, sequence are as follows: 27F:5 '-AGAGTTTGATCCTGGCTCAG-3 '；1492R:5'-TACGGCTACCTTACGACTT-
3’。
PCR response procedures are as follows: 95 DEG C of 5min；95 DEG C of 30s, 56 DEG C of 90s, 72 DEG C of 60s (33cycles)；72℃10min；4
℃。
After extension increasing sequence carries out purifying sequencing, tetraploid rice is carried out by the Blastn program of NCBI, it is soft using MEGA5
Part establishes phylogenetic tree with Neighbor-Joining method.
Three, the biological characteristics of bacterial strain
It carries out 94 kinds of phenotypes to Streptomycesalbidoflhaving W68 using Biolog GEN III microwell plate to test, including 71 kinds of carbon
(table 2, table 3) is tested using test and 23 kinds of chemosensitivities in source.
Utilization power of the 2. Streptomycesalbidoflhaving W68 of table to 71 kinds of different carbon sources
Remarks :+indicate can use ,-indicate to utilize
Tolerance situation of the 3. Streptomycesalbidoflhaving W68 of table to 23 kinds of different reagents
Remarks :+indicate tolerance ,-indicate not tolerating
Four, the preservation of bacterial strain
The Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge is preserved in Chinese microorganism strain on March 14th, 2016
Preservation administration committee common micro-organisms center (abbreviation CGMCC, address are as follows: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3),
Deposit number is CGMCC No.12210.
Embodiment 2
It is isolated from the antifungal activity test of the Streptomycesalbidoflhaving W68 of deep-sea sludge
Using the antifungal activity of tablet face-off method measurement Streptomycesalbidoflhaving W68:
(1), streptomycete culture and collection: flat in PDA using method of scoring inoculation Streptomycesalbidoflhaving Streptomycesalbidoflhaving W68
On plate, 28 DEG C culture 5-7 days to generating enough spores；It is spare that spore suspension is made in scraping spore in appropriate amounts of sterilized water.
(2), the culture of pathogen: inoculation Rhizoctonia solani, cabbage oxysporum, Cochliobolus sativus, Pyricularia oryzae,
In on PDA plate, 28 DEG C of cultures use after a week for cotton-wilt fusarium, verticillium dahliae.
(3), it plate face-off experiment: is inoculated with Bacteria culturing object fungus block respectively in PDA plate center, apart from pathogen
2.0cm or so is inoculated with 5 μ l of Streptomycesalbidoflhaving W68 spore suspension, and 28 DEG C are cultivated 3-5 days, and growth of pathogenic bacteria situation is observed.
Embodiment 3
Streptomycesalbidoflhaving W68 is the microbial bacterial agent of material:
(1) preparation of liquid seeds
Picking grows aerial hyphae and the spore of 4-5 days Streptomycesalbidoflhaving W68 on PDA plate, is seeded to 2 × YT
In fluid nutrient medium, 28-30 DEG C, 180rpm shaken cultivation is used after 1-2 days as seed liquor.
(2) preparation of solid fermentation seed
By the access of liquid seeds in mass ratio 10% equipped in the tissue culture bottle of solid fermentation material, 28-30 DEG C, 4-6 is cultivated
It, obtains solid fermentation seed.
(3) microbial bacterial agent
With agricultural wastes feces of livestock and poultry, edible fungi residue, wheat bran or dregs of beans etc. for base stock, add a certain proportion of
MnSO4、(NH)2SO4, material-water ratio 1:1.Solid fermentation seed is inoculated with according to 10% ratio.28-30 DEG C culture 5-7 days, fermentation
It is air-dried after culture, spore concentration can achieve 1 × 1010A/g is used as microbial streptomycete microbial inoculum.
Embodiment 4
Streptomycesalbidoflhaving W68 is prepared by the microbial manure of material:
(1) preparation of liquid seeds
Picking grows aerial hyphae and the spore of 4-5 days Streptomycesalbidoflhaving W68 on PDA plate, is seeded to 2 × YT
In fluid nutrient medium, 28-30 DEG C, 180rpm shaken cultivation is used after 1-2 days as seed liquor.
(2) preparation of solid fermentation seed
By the access of liquid seeds in mass ratio 10% equipped in the tissue culture bottle of solid fermentation material, 28-30 DEG C, 4-6 is cultivated
It, obtains solid fermentation seed.
(3) preparation of microbial manure
With agricultural wastes feces of livestock and poultry, edible fungi residue, wheat bran or dregs of beans etc. for base stock, add a certain proportion of
MnSO4、(NH)2SO4, material-water ratio 1:1.Solid fermentation seed is inoculated with according to 10% ratio.28-30 DEG C culture 5-7 days, fermentation
It is air-dried after culture, spore concentration can achieve 1 × 1010A/g, uses as microbial streptomycete fertilizer.
Embodiment 5
It is isolated from the microbial bacterial agent or Micro-biological Fertilizer on Plant fungal disease of the Streptomycesalbidoflhaving W68 of deep-sea sludge
Prevention and control application:
One, prevention and control of the Streptomycesalbidoflhaving W68 of deep-sea sludge to root rotof flax are isolated from
(1) test site and environmental condition: Institute of Microorganism, Academia Sinica's outdoor elements carry out three times respectively
Pot experiment.
(2) test medicine: blank control, Streptomycesalbidoflhaving W68 spore suspension, 70% first support pulvis
(3) test method:
Seed treatment: taking wheat seed several, aseptic water washing 2 times, then sterile after 10%84 thimerosals handle 5min
Water logging kind 4h；
Seed coating is carried out with Streptomycesalbidoflhaving W68: taking soaked seed in culture dish, uses 1ml carboxylic respectively
The W68 spore suspension that methylcellulose sodium solution and 1ml are gathered carries out seed coating, stands overnight for 28 DEG C after mixing；
Bored kind of seed is carried out with first support: according to the application method of first support, it is bored that 6h being carried out to the wheat seed of surface sterilization
Kind processing.
(4) processing arrangement: each flowerpot plants 10 seeds, and four parallel, random alignments are done in each processing, outdoor natural
Growing state is observed in environmental condition culture 2-3 weeks.The experiment is in triplicate.
(5) statistical data: the diameter of the height of plant, the weight in wet base of overground part plant and dry weight, plant haulm is remembered
Record, statistical data analysis.
(6) interpretation of result: in the pot experiment prevented and treated root rotof flax, using the coated method of seed, use is micro-
White yellow streptomycete W68 bacteria preparation improves emergence rate of the wheat seed in the soil containing Cochliobolus sativus, plant height, plant
Dry weight is above the control of morbidity group, and can achieve level of growth (Fig. 3) of the wheat seed in no pathogen soil substantially.
Two, it is isolated from field experiment of the Streptomycesalbidoflhaving W68 to Muskmelon Fusarium wilt of deep-sea sludge.
(1) test site and environmental condition: the experiment is located at Xinzhou City Donglou township, Shanxi Province.Experimental plot middle fertility, it is preceding
Stubble crop is muskmelon.Muskmelon sowing, cultivation, management part uniformity, meets local agricultural practice.
(2) demonstrate medicament: blank control, Streptomycesalbidoflhaving W68 be material microbial bacterial agent or microbial manure, its
Its company's similar product.
(3) plot area and processing arrange: each processing 70m2, with covering with ground sheeting, 2 row muskmelons are planted in each processing.
(4) application method: seed soaking method is used, is impregnated 1 hour before muskmelon sowing with bacterial manure, will after planting steep seed
Bacterium solution tank in kind of cave.
(5) data statistics: taking pictures, and checks dead seedling, statistical results.
(6) interpretation of result: being micro- life of material using Streptomycesalbidoflhaving W68 by the seed soaking to muskmelon seeds
Object microbial inoculum or microbial manure can significantly overcome the muskmelon continuous cropping obstacle as caused by fusarium wilt: cross muskmelon in last year kind
Field continuously plant muskmelon, the muskmelon shoot survival percent of processing group can achieve 93%, the muskmelon shoot survival percent of blank control group
Only 13% (Fig. 4).
Three, it is isolated from field experiment of the Streptomycesalbidoflhaving W68 to phytoph-thora capsici leonian of deep-sea sludge.
(1) test site and environmental condition: the experiment is located at the village Xinzhou City Gao Cheng, Shanxi Province.Preceding crop is capsicum, product
Kind is that Beijing is red.Pepper seedling raising.
(2) demonstrate medicament: blank control, Streptomycesalbidoflhaving W68 be material microbial bacterial agent or microbial manure, its
His company's similar product.
(3) plot area and processing arrange: each processing 70m2, with covering with ground sheeting, 2 row capsicums are planted in each processing.
(4) application method: being handled 2-3 hours when capsicum transplantation of seedlings with bacterial manure root dipping, fills bacterium solution in kind of cave after transplanting.
(5) data statistics: taking pictures, and carries out output statistics.
(6) interpretation of result: pepper seedling uses Streptomycesalbidoflhaving W68 for the microbial bacterial agent of material or micro- life when transplanting
Object fertilizer carries out root dipping processing to seedling, can significantly mitigate the occurring degree of phytoph-thora capsici leonian.Compared with the control group, root dipping is handled
The plant strain growth cycle stretch-out crossed about 1-2 weeks, and chilli yield increases by 5% (Fig. 5).
Four, it is isolated from field experiment of the Streptomycesalbidoflhaving W68 to take-all of deep-sea sludge.
(1) test site and environmental condition: the test is located at Hebei province, overflow river town, the Jing County village Dong Li village.Preceding crop is
Corn, the long-term per unit area yield of wheat are averaged 950 jin or so per acre.Wheat cultivation, 30 pounds/acre.The rich board fertilizer specially for wheat in bottom application field per acre
100 jin, it is grab moisture sowing when irrigating situation sowing, jelly is poured after broadcasting water 1 time, pour water of turning green, mu applies 60 jin of urea.With 10% benzene sulphur
Grand+56% first tetrachloro sodium controlling weeds prevent and treat wheat midge, aphid with 25g/L gamma cyhalothrin.
(2) it demonstrates medicament: blank control, the microbial bacterial agent that Streptomycesalbidoflhaving W68 is material or microbial manure, another
If chemical agent compares the 50% triazolone WP and net FS of 12.5% total eclipse.
(3) plot area and processing arrange: per kilogram wheat seed uses 50g bacterial manure.
(4) data statistics: investigation emergence rate, wheat during jointing stage and heading stage respectively carry out a root system investigation, take diagonal
5 point sampling of line, every takes 20 plants, and investigation root system is infected situation, by take-all grade scale record, statistics diseased plant rate
And disease index.Dead ears are investigated, take 1 meter of duplicate rows, investigates total spike number and dead ears number, counts dead ears rate at every.Survey and produce investigation,
Each random 5 points of samplings of processing, investigate 1 meter of total spike number of duplicate rows, count mu spike number, and every takes 20 fringe of number of productive ear, threshing after doing,
Grain number per spike and mass of 1000 kernel are counted, yield and effect of increasing production are calculated.
(5) interpretation of result: deep-sea Streptomycesalbidoflhaving W68 is that the use of the microbial bacterial agent or microbial manure of material can
To significantly reduce the diseased plant rate of wheat heading stage, disease index is reduced.Compared with chemical agent, Streptomycesalbidoflhaving W68 is material
The microbial bacterial agent of material or the preventive effect of microbial manure are substantially better than chemical agent triazolone, suitable with the net preventive effect of total eclipse.It can
See the use that can substitute chemical agent completely using Streptomycesalbidoflhaving W68 as the microbial bacterial agent of material or microbial manure
(Fig. 6).
Claims (6)
1. one plant of Streptomycesalbidoflhaving (Streptomyces albidoflavus) bacterial strain W68, deposit number CGMCC
No.12210。
2. a kind of microbial bacterial agent, it is characterised in that: its active constituent is Streptomycesalbidoflhaving bacterial strain described in claim 1
W68。
3. the application of microbial bacterial agent described in claim 2, it is characterised in that: the microbial bacterial agent is for preventing and treating soil-borne fungus
Disease, the soil-borne fungus disease are by following a kind of fungi or a variety of fungus-caused: fusarium verticillioides, sharp spore reaping hook
Bacterium, Bipolaris sacchari, Rhizoctonia solani Kuhn, Verticillium dahliae bacterium, grey big angle base shell bacterium, every gram of dry weight of the microbial bacterial agent
Contain 1 × 10 containing the Streptomycesalbidoflhaving W68 for being isolated from deep-sea sludge10A spore.
4. the application of microbial bacterial agent described in claim 2, which is characterized in that the microbial bacterial agent is used as microbial manure.
5. the application of microbial bacterial agent as claimed in claim 3 is used to prevent and treat root rotof flax, Muskmelon Fusarium wilt, capsicum epidemic disease
Mildew and take-all.
6. a kind of preparation method of microbial manure, this method comprises:
(1) preparation of liquid seeds
Picking grows aerial hyphae and the spore of 4-5 days Streptomycesalbidoflhaving W68 as described in claim 1 on PDA plate,
It is seeded in 2 × YT fluid nutrient medium, 28-30 DEG C, 180rpm shaken cultivation uses after 1-2 days as seed liquor；
(2) preparation of solid fermentation seed
By the access of liquid seeds in mass ratio 10% equipped in the tissue culture bottle of solid fermentation material, 28-30 DEG C, cultivates 4-6 days, obtain
To solid fermentation seed；
(3) preparation of microbial manure
Using agricultural wastes feces of livestock and poultry, edible fungi residue, wheat bran or dregs of beans as raw material, a certain proportion of MnSO is added4、(NH)2SO4, material-water ratio 1:1 is inoculated with solid fermentation seed according to 10% ratio, 28-30 DEG C culture 5-7 days, after fermented and cultured
It air-dries, spore concentration reaches 1 × 1010A/g, uses as microbial streptomycete fertilizer.
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