CN106635878A - Streptomyces sp. 5X4 and applications thereof as biocontrol bacterium in preventing and treating plant diseases - Google Patents
Streptomyces sp. 5X4 and applications thereof as biocontrol bacterium in preventing and treating plant diseases Download PDFInfo
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- CN106635878A CN106635878A CN201610934021.XA CN201610934021A CN106635878A CN 106635878 A CN106635878 A CN 106635878A CN 201610934021 A CN201610934021 A CN 201610934021A CN 106635878 A CN106635878 A CN 106635878A
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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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- 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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Abstract
The invention relates to the biological prevention and treatment for plant diseases, belongs to the fields of biological prevention and treatment research and applications in the subject of plant protection, and in particular relates to Streptomyces sp. 5X4 and applications of Streptomyces sp. 5X4 as a biocontrol bacterium in treating plant diseases. The Streptomyces sp. 5X4 strain is assigned with the accession number of CGMCC No.12605. The strain can effectively inhibit the growth of plant pathogenic fungi including vegetable botrytis cinera, phomopsis amygdali, monilia fructigena, sclerotinia sclerotiorum, fusarium wilt of cucumber and the like. The prevention and treatment effects of Streptomyces sp. 5X4 for grey mould fruit rot of strawberry in the field are obviously superior to those of the conventional chemical agents including pyrimethanil and procymidone, the prevention effects are improved by about 25% and 30% respectively, and the Streptomyces sp. 5X4 strain has the potential of preventing and treating various plant diseases. The increasing of the biological agent and the reducing of chemical agents are beneficial for protecting the environment, reducing the pesticide residue and promoting the agricultural sustainable development.
Description
Technical field
The present invention relates to the biological control of plant disease, belongs to the biological control research and application neck in plant protection subject
Domain.
Background technology
Plant disease is to restrict plant one of major obstacle of growth safely, and current prevention and controls mainly spray chemical drugs
Agent.Because environmental condition is suitable, protecting field crop pest occurs seriously, frequently there is the resistance to the action of a drug using chemical agent and agricultural chemicals is residual
Equivalent risk is stayed, is developed and can effectively be reduced the usage amount of chemical agent using biological pesticide.There is antagonism to make phytopathogen
Microorganism is the main source for developing biological pesticide, and this kind of microbe species are various, widely distributed, even if same micro- life
, due to being distributed in different regions, its antagonism is strong and weak or mechanism also has different for thing.Biocontrol microorganism resource is excavated, first
It is extensively to screen Antagonistic Fungi with targeted pathogens, this is the element task in biological control research field.
Include Antagonism power, antimicrobial spectrum to the basic research of Antagonistic Fungi, the stability of antagonism performance, antagonism, resist
Bacterium substance classes and metabolism etc..Developing biological pesticide includes fermentation condition optimization, auxiliary agent use, medicament development of Antagonistic Fungi etc..
Biological pesticide Field information includes optimum time, prevention and controls, dosage and number of times of controlling disease etc..It is all these all
Must set up on the basis of the preferable Biocontrol microorganism bacterial strain of antagonism performance is obtained.
The content of the invention
It is an object of the invention to provide the preferable Biocontrol microorganism bacterial strain of one plant of antagonism performance, for preventing and treating protecting field crop
Disease chemical agent usage amount greatly, often, easily causes the resistance to the action of a drug and Pesticide Residue, should reduce making for chemical agent
With enhancing bio-control method.Screening obtains streptomycete 5X4 bacterial strains, premenstruum (premenstrua) Primary Study, 5X4 from soil in the present invention
Ability with stronger antagonistic plant pathogenic fungi.To its further research, the possibility with research and development novel biopesticide.
The invention discloses a kind of streptomycete (Streptomyces sp.) 5X4, its bacterial strain deposit number is CGMCC
No.12605。
It can suppress various plants disease fungus to grow to streptomycete 5X4 of the present invention, can be used for controlling plant diseases
The purposes of biocontrol bacteria.
Further, the plant disease is by ash arrhizus bacteria, peach branch-rot bacterium, Monilinia fructicola, Sclerotinia sclerotiorum
Cause with cucumber fusarium axysporum.
5X4 bacterial strains are, with Botrytis cinerea as targeted fungal, to screen from soil and obtain.Jing molecular biology methods are accredited as
Streptomycete (Streptomyces sp.).5X4 can effectively suppress the growth of Botrytis cinerea germ (Botrytis cinerea), right
Other germs such as peach branch-rot bacterium (Phomopsis amygdali), Monilinia fructicola (Monilinia fructicola), oil
Dish hyphal cluster germ (Sclerotinia sclerotiorum) and cucumber fusarium axysporum (Fusarium oxysporum
) etc. f.sp.cucumerinum also there is preferable antagonism.Determine after shifting for 20 generations, its antagonism stable performance.Prevent in field
Control grey mould fruit rot of strawberry test and show that the tunning of the bacterial strain has preferable preventive and therapeutic effect to grey mould fruit rot of strawberry, and preventive effect is substantially excellent
In chemical agent pyrimethanil and Sukeling, prevention effect has been respectively increased 25% and 30% or so.
Description of the drawings
Fig. 1 is antagonistic effects of the streptomycete 5X4 to Botrytis cinerea germ；
Fig. 2 is inhibitory action of the streptomycete 5X4 tunnings to Botrytis cinerea germ under different incubation times；
Fig. 3 applies Field strawberries gray mold after streptomycete 5X4 fermentate 10d for the 2nd time, and a situation arises；
Fig. 4 is phylogenetic evolution trees of the actinomyces 5X4 based on 16S rDNA.
It is general that 5X4 bacterial strains of the present invention were stored in China Committee for Culture Collection of Microorganisms on June 12nd, 2016
Logical microorganism center, (Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica), Classification And Nomenclature is
Streptomycete (Streptomyces sp.), deposit number is CGMCC No.12605.
Specific embodiment
1st, strain isolation
(1) different regions soil sample 1g is taken respectively, add water and make certain density Soil Slurry, then carry out gradient dilution
103～105Times, 100 μ l dilution coated plates are taken, put in 28 DEG C of incubators and cultivate 48h；
(2) further line culture, purifying, and picking single bacterium colony preserve standby to picking single bacterium colony on Gause I culture medium
With.
2nd, antagonistic activity is determined
(1) with Botrytis cinerea germ as targeted fungal, the Antagonism of isolated strains, 24 DEG C of trainings are determined using opposite culture method
4～5d is supported, antibacterial bandwidth is determined.174 plants of actinomyces are separated to obtain from soil altogether, wherein 7 plants have by force to Botrytis cinerea germ
Antagonism (antibacterial band >=1.00cm), the antibacterial bandwidth of actinomyces 5X4 reaches 1.50cm (table 1, Fig. 1).
Table 1 has the actinomycetes strain of strong antagonistic activity to Botrytis cinerea germ
(2) the bacterial strain 5X4 of the most strong antagonistic activity of tool is chosen, same method determines its antagonism spectrum, finds it to peach branch-rot
The plant pathogenic fungis such as bacterium, Monilinia fructicola, Sclerotinia sclerotiorum and cucumber fusarium axysporum have stronger antagonism (table 2)；
Antagonisms of the actinomyces 5X4 of table 2 to different plant pathogenic fungis
(3) bacterial strain squamous subculture 5,10,15 and after 20 generations on Gause I culture medium will be obtained, will be examined by opposite culture
Its stability to aimed strain antagonism is surveyed, is as a result shown, turned culture and have no effect on its antagonistic ability, generation of tube 5,10
After generation, 15 generations and 20 generations, its antibacterial bandwidth to Botrytis cinerea germ is respectively 1.50,1.45,1.53 and 1.47cm；
(4) the 5X4 ferment filtrates of culture 1,2,4 and 6d are taken respectively, is added in Gause I culture medium (V/V=1/5), survey
Fixed its inhibitory action to Botrytis cinerea pathogen growth, optimal with the zymotic fluid fungistatic effect of 28 DEG C of culture 4d, inhibiting rate reaches
76.30% (Fig. 2)；
3rd, field efficacy is determined
(1) by test strain Gause I nutrient solution concussion and cultivate 4d, unwrapping wire bacterium suspension is obtained；
(2) the strawberry growth field that fruit has been fallen ill is selected, all disease fruits are removed, bacterium solution is sprayed in fruit surface；
(3) compareed as chemical agent with preventing and treating the conventional medicament Sukeling of grey mould fruit rot of strawberry and pyrimethanil, clear water is blank right
According to.Dispenser 2 times altogether, is spaced 10d.Per cell 20m2, 3 repetitions.
After (4) the 2nd dispenser 10d, the order of severity of dispenser cell grey mould fruit rot of strawberry substantially mitigates compared with the control (figure
3), the field efficacy of 5X4 culture suspensions distinguishes high by 25% and 30% up to 69.11% than the preventive effect of pyrimethanil and Sukeling
Left and right (table 3).
Field efficacies of the actinomyces 5X4 of table 3 to grey mould fruit rot of strawberry
4th, bacterial strain Identification of Species
(1) CTAB methods extract bacterial genomes；
(2) with extract genomic DNA as template, with 16S rDNA universal primer 16s_63f and 16s_1387 as upper
Downstream primer enters performing PCR amplification, and amplified production is sequenced；
(3) sequence for measuring is compared into analysis in GenBank, using Mega software building phylogenetic evolution trees, mirror
It is streptomycete (Fig. 4) to determine it.
Claims (3)
1. streptomycete (Streptomyces sp.) 5X4, the bacterial strain deposit number is CGMCC No.12605.
2. use of (Streptomyces sp.) 5X4 of streptomycete described in claim 1 as the biocontrol bacteria of controlling plant diseases
On the way.
3. purposes as claimed in claim 2, it is characterised in that the plant disease is by ash arrhizus bacteria, peach branch-rot bacterium, peach
What brown rot germ, Sclerotinia sclerotiorum and cucumber fusarium axysporum caused.
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| CN201610934021.XA CN106635878A (en) | 2016-10-25 | 2016-10-25 | Streptomyces sp. 5X4 and applications thereof as biocontrol bacterium in preventing and treating plant diseases |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112646757A (en) * | 2021-01-25 | 2021-04-13 | 浙江师范大学 | Streptomyces syringae and application thereof in plant disease control |
| CN112725250A (en) * | 2021-03-10 | 2021-04-30 | 扬州大学 | Streptomyces lavendulae strain 2-1-2F-1 and application of biomass thereof in prevention and treatment of vegetable soil-borne diseases |
| CN114214236A (en) * | 2021-12-22 | 2022-03-22 | 湖南省农业生物技术研究所 | Streptomyces scorpion LSS-F01 and application thereof in prevention and treatment of fungal diseases of crops |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112646757A (en) * | 2021-01-25 | 2021-04-13 | 浙江师范大学 | Streptomyces syringae and application thereof in plant disease control |
| CN112646757B (en) * | 2021-01-25 | 2022-08-26 | 浙江师范大学 | Streptomyces syringae and application thereof in plant disease control |
| CN112725250A (en) * | 2021-03-10 | 2021-04-30 | 扬州大学 | Streptomyces lavendulae strain 2-1-2F-1 and application of biomass thereof in prevention and treatment of vegetable soil-borne diseases |
| CN114214236A (en) * | 2021-12-22 | 2022-03-22 | 湖南省农业生物技术研究所 | Streptomyces scorpion LSS-F01 and application thereof in prevention and treatment of fungal diseases of crops |
| CN114214236B (en) * | 2021-12-22 | 2023-08-18 | 湖南省农业生物技术研究所 | Streptomyces scorpion LSS-F01 and application thereof in crop fungal disease control |
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