CN108048380A - One plant of Deccan streptomycete QY-3 and its application - Google Patents
One plant of Deccan streptomycete QY-3 and its application Download PDFInfo
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Abstract
The invention discloses one plant of Deccan streptomycete QY 3 and its applications, the entitled Streptomyces deccanensis of Latin of the Deccan streptomycete, bacterial strain number is QY 3, is preserved in China General Microbiological culture presevation administrative center, and deposit number is CGMCC No.14467.Deccan streptomycete QY 3 of the present invention has the anti-fungal activity of plant pathogenic of wide spectrum, the phytopathogens such as germ, Botryosphaeria berengeriana f. sp, mango blossom-end rot bacterium or guava plan disk stey that fester to colletotrichum gloeosporioides Penz, Glorosprium musarum Cookeet Mass, banana blight bacteria, fusarium graminearum, cucumber fusarium axysporum, dragon fruit sickle-like bacteria, dragon fruit have obvious inhibiting effect, especially rubber tree colletotrichum gloeosporioides Penz has good control effect, is the biocontrol actinomycetes strain of one plant of great exploitation potential.
Description
Technical field
The present invention relates to biocontrol of plant disease fields, and in particular to one plant of Deccan streptomycete QY-3 and its application.
Background technology
Rubber tree (Hevea brasiliensis) is perennial tropical rain forest arbor species, and sea is distributed mainly in China
The areas such as south, Yunnan, Guangdong, Guangxi, Fujian are the main sources of natural rubber.Natural rubber be the important raw material of industry and
Strategic resources, the daily life, national defence and industrial construction with the people are closely bound up, and superior performance can't be closed at present
Substitute completely into rubber, occupy an important strategic position in national economy.Rubber anthracnose is endanger China rubber tree one
The important leaf diseases of kind, are one of four major diseases in current rubber production, it is straight which can endanger rubber seedling, crop field treelet
To into age tapped tree, it can cause the fallen leaves repeatedly of rubber tree and tender tip time are withered, and postponement is opened to cut the time when serious, under gum yield is notable
Drop.Rubber anthracnose is very big to the rubber harm in the area such as China Hainan, Yunnan, Guangdong and threatens the production of China's natural rubber
One of important biomolecule factor that industry is lasting, develops in a healthy way.
Colletotrichum gloeosporioides Penz [Colletotrichum gloeosporioides (Penz.) Penz.And Sacc.] is to cause
A kind of important pathogen of anthracnose of rubber trees is mainly prevented at present for the prevention of the pathogen using agricultural measures and chemistry
It controls, it is direct using bactericidal agent for preventing and treating effect, but easily develop immunity to drugs, breeding resistant variety needs to put into a large amount of human and material resources
And the time, and varietal resistance is also easier to lose.Therefore research safety effectively preventing measure is imperative, and wherein biological control is
Most there is one of measure of application prospect.Actinomyces are the quasi-microorganisms for having huge practical value, are sent out at present from microorganism
In about 8000 existing various bioactivators, nearly 70% is generated by actinomyces.Actinomyces are the weights of farm antibiotics
Want source, using actinomyces generate secondary metabolite prepare microbial pesticide, because its with it is pollution-free, be not easy to make to be harmful to
The features such as biology develops immunity to drugs, it has also become the main body of public nuisance-free agricultural chemicals and the developing direction of pesticide in future.
Part work, Choudhary etc. has also been made in terms of the screening of anthrax-bacilus biocontrol actinomycetes both at home and abroad from India's sheet
Soil is separated to one plant of biocontrol actinomycetes MT7, grey violet streptomycete (Streptomyces violascens) is accredited as, to a variety of water
The postharvest pathogenic fungi of fruit has the fungistatic effect of wide spectrum, is such as to the minimum inhibitory concentration (MIC) of colletotrichum gloeosporioides Penz
0.0312mg/mL；The fermentation broth coarse extract of the S.malaysiensis MJM1968 of the discoveries such as Palaniyandi is to Chinese yam anthrax
Germ has compared with high inhibition effect, and 50 μ g/mL fermentation broth coarse extracts can completely inhibit colletotrichum gloeosporioides Penz conidia germination；Du Yi
Separation screening is to one plant of actinomyces S15 in the pedotheque gathered from Jinggang Mountains such as new, and the bacterium culturing filtrate is to big
A variety of important plant pathogenic fungis such as beans anthrax bacteria, citrus anthracnose bacterium, cucumber anthracnose have stronger inhibition to make
With antagonism spectrum is wider, is accredited as streptomyces bottropensis (S.bottropensis)；Li Juan etc. is separated to one from rhizosphere of strawberry soil
Strain actinomyces DTJ-24, reaches 69.89% to the inhibiting rate of strawberry colletotrichum gloeosporioides Penz, is accredited as Strepiomyces lavendulae
(S.lavendulae).The biocontrol actinomycetes of the anthrax-bacilus screened at present are for Chinese yam, strawberry, citrus, grape, soybean mostly
Crops are waited, and the research that biocontrol actinomycetes screening is carried out for rubber anthrax bacteria is rarely reported.
The content of the invention
For in place of more than the deficiencies in the prior art, the present invention provides one plant of Deccan streptomycete QY-3 and its application.
The technical solution that the present invention takes is as follows：
One plant of Deccan streptomycete QY-3, the entitled Streptomyces deccanensis of Latin of the Deccan streptomycete,
Bacterial strain number is QY-3, is preserved in China General Microbiological culture presevation administrative center, and deposit number is CGMCC No.14467, is protected
Hide address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3.
Preferably, the Deccan streptomycete QY-3 is used to inhibit plant pathogenic fungi.
Preferably, the plant pathogenic fungi include colletotrichum pathogen, banana blight bacteria, fusarium graminearum,
Fester germ, Botryosphaeria berengeriana f. sp, mango blossom-end rot bacterium or guava of cucumber fusarium axysporum, dragon fruit sickle-like bacteria, dragon fruit is intended
Disk stey.
It is furthermore preferred that the colletotrichum pathogen is colletotrichum gloeosporioides Penz or Glorosprium musarum Cookeet Mass.
The present invention also provides a kind of zymotic fluid, the zymotic fluid is prepared by the Deccan streptomycete QY-3 of the present invention.
Preferably, the zymotic fluid is prepared by the following：Fermented and cultured is carried out to Deccan streptomycete QY-3, crosses and filters out
Thalline is gone to obtain zymotic fluid；Fermentation medium is used by fermented and cultured：Millet 20g, peptone 3g, glucose sugar 20g,
CaCO32g, NaCl 2.5g, water 1000mL, pH7.2-7.4, condition of culture is 28 DEG C, 180rpm shaken cultivations 7d.
The present invention also provides the method for screening and separating of the Deccan streptomycete QY-35, comprise the following steps：
Glacial deposit sample is taken, grinds, is added in the container of built-in sterile water and bead, vibrates, takes supernatant
It is diluted, obtains dilution, dilution is taken to be coated on Gao Shi I culture medium flat plates uniformly, cultivates 5-10d, picking actinomyces
Bacterium colony is further isolated and purified using method of scoring, until obtaining actinomyces strain pure culture；Using colletotrichum gloeosporioides Penz as target bacterium, adopt
With opposite culture method, one plant of bacterial strain QY-3 to colletotrichum gloeosporioides Penz with higher inhibitory activity is screened.
Preferably, the method for screening and separating of the Deccan streptomycete QY-3, it is isolated by the following method：
10g glacial deposit samples are taken, fully grinds, adds in the container of built-in 100mL sterile waters and bead, in shaking
100rpm vibrates 20min on bed, and supernatant is taken to carry out 10 times of gradient dilutions, obtains dilution, takes dilution 0.1mL in Gao Shi I
Be coated with uniform on culture medium flat plate, be placed in 28 DEG C of constant incubators and cultivate 5-10d, picking actinomyces bacterium colony using method of scoring into
One step isolates and purifies, until obtaining actinomyces strain pure culture；Using colletotrichum gloeosporioides Penz as target bacterium, using opposite culture method, sieve
Choose one plant of bacterial strain QY-3 to colletotrichum gloeosporioides Penz with higher inhibitory activity.
Compared with prior art, the beneficial effects of the invention are as follows：
Deccan streptomycete QY-3 of the present invention has the anti-fungal activity of plant pathogenic of wide spectrum, to colletotrichum gloeosporioides Penz, banana charcoal
Subcutaneous ulcer germ, banana blight bacteria, fusarium graminearum, cucumber fusarium axysporum, dragon fruit sickle-like bacteria, dragon fruit fester germ, apple
Fruit Target spot pathogen, mango blossom-end rot bacterium and guava, which intend the phytopathogens such as disk stey, has obvious inhibiting effect, especially
Rubber tree colletotrichum gloeosporioides Penz has good control effect, is the biocontrol actinomycetes strain of one plant of great exploitation potential.
Description of the drawings
Fig. 1 is Deccan streptomycete QY-3 colonial morphologies (A) and spore filament shapes (B)；
Fig. 2 is Deccan streptomycete QY-3 to antagonistic activity in the ware of colletotrichum gloeosporioides Penz；
Fig. 3 is the Stability Determination result of Deccan streptomycete QY-3 zymotic fluids；
Fig. 4 is control effect of the Deccan streptomycete QY-3 zymotic fluids to sense anthracnose rubber tree leaf.
Specific embodiment
The present invention is described in further detail below by specific embodiment combination attached drawing.
Experimental method used in the embodiment of the present invention is conventional method unless otherwise specified.
Material, reagent used in the embodiment of the present invention etc., are commercially available unless otherwise specified.
The separation screening of 1 Deccan streptomycete of embodiment (Streptomyces deccanensis) QY-3
Glacier topsoil is taken to take 10g glacial deposit samples to get glacial deposit, fully grind, add in built-in
In the triangular flask of 100mL sterile waters and bead, 20min is vibrated in 100rpm on shaking table, supernatant is taken to carry out 10 times of gradients dilute
It releases, obtains dilution, dilution 0.1mL is taken to be coated on Gao Shi I culture medium flat plates uniformly, is placed in 28 DEG C of constant incubators and trains
5-10d is supported, picking actinomyces bacterium colony is further isolated and purified using method of scoring, until obtaining actinomyces strain pure culture；With glue
Spore anthrax-bacilus are target bacterium, using opposite culture method, screen one plant of bacterial strain to colletotrichum gloeosporioides Penz with higher inhibitory activity
QY-3。
The biological property of Deccan streptomycete (Streptomyces deccanensis) QY-3 of the present invention is as follows：
The 16S rDNA sequences (see sequence table) of Deccan streptomycete QY-3 effectively fragment length is 1449bp, is submitted
GenBank database logins number are：MG751325.Effective kind of sequence similarity search is carried out in EzTaxon databases, is sent out
Existing Deccan streptomycete QY-3 and the bacterial strain of streptomyces are highly relevant, are the actinomyces of streptomyces, and Deccan streptomycete QY-
3 streptomycetes and the similitude highest of Streptomyces deccanensis DAS-139 (T), similarity 99.65%, with
The similarity of Streptomyces scabieiNRRL B-16523 (T) is 99.59%, with Streptomyces
The similarity of europaeiscabiei KACC 20186 (T) is 99.52%, is existed with the similarity of other streptomyces
Between 97.83-99.00%.
Deccan streptomycete QY-3 28 DEG C of culture 7d on No. I tablet of Gao Shi observe its colonial morphology as circle, flat, in vain
Color, is with a loop pile hair, micro- fold outside, and spore filament shapes are that spiral two-stage is verticillate (Fig. 1).
Deccan streptomycete QY-3 is inoculated in different solid mediums, and 28 DEG C of culture 7-15d observe its growth situation, cultivate
Feature is as shown in table 1.
The cultural characteristic of 1 Deccan streptomycete QY-3 of table
The biochemical reactions and utilization of carbon source situation of Deccan streptomycete QY-3 are measured respectively, the results are shown in Table 2 and table 3.Knot
Fruit shows that Deccan streptomycete QY-3 can be with hydrolysis starch, and liquefy gelatin, it is not possible to using cellulose, can reduce nitrate, energy
Milk generation is enough made to peptonize phenomenon and solidify, not chromogenesis and H2S (table 2).In terms of utilization of carbon source, it can utilize Portugal
Grape sugar, raffinose, D- galactolipins, D-Fructose, rhamnose, it is not possible to utilize arabinose, mannitol, inositol, sucrose, malt
Sugar, D- xyloses (table 3).
The biochemical reactions of 2 Deccan streptomycete QY-3 of table
Note："+" is the positive, and "-" is feminine gender
3 Deccan streptomycete QY-3 utilization of carbon source situations of table
Note："+" is the positive, and "-" is feminine gender
Embodiment 2：Deccan streptomycete QY-3 measures the inhibitory activity of pathogen
Fermentation medium used in the present invention：Millet 20g, peptone 3g, glucose sugar 20g, CaCO32g, NaCl
2.5g, water 1000mL, pH：7.2-7.4, condition of culture are 28 DEG C, 180rpm shaken cultivations 7d.
Inhibitory activity of the Deccan streptomycete QY-3 to colletotrichum gloeosporioides Penz is determined using tablet dual test, such as Fig. 2 institutes
Show, Deccan streptomycete QY-3 has stronger antagonistic activity to colletotrichum gloeosporioides Penz.
Fermented and cultured is carried out to Deccan streptomycete QY-3, thalline is filtered to remove and obtains fermented liquid supernatant, test it to not
With the inhibitory activity of plant pathogenic fungi.By table 4 as it can be seen that the zymotic fluid of Deccan streptomycete QY-3 has the antimycotic work of wide spectrum
Property, particularly there is stronger inhibitory activity, such as suppression to colletotrichum gloeosporioides Penz and Glorosprium musarum Cookeet Mass to colletotrichum pathogen
System activity is respectively 87.0% and 88.9%.
4 Deccan streptomycete QY-3 of table is to the inhibitory activity of different plant pathogenic fungis
Embodiment 3：The temperature and pH Stability Determinations of Deccan streptomycete QY-3 zymotic fluids
Deccan streptomycete QY-3 zymotic fluids are placed under different temperatures and different pH and handle 1h, measure it to colletotrichum gloeosporioides Penz
Inhibitory activity.As shown in Figure 3, Deccan streptomycete QY-3 zymotic fluids have preferable temperature and pH stability.
Embodiment 4：Deccan streptomycete QY-3 zymotic fluids are to the control effect of sense anthracnose rubber tree leaf
Freshly harvested rubber tree blade is respectively placed in sterile water, Bravo (1000 μ g/mL), bacterial strain QY-3 zymotic fluids
Middle immersion 1h takes out placement shady place and dries, and using sterilizing, syringe needle manufactures wound on blade, and glue spore anthrax is inoculated in wound
Bacterium bacteria cake, moisturizing culture 3-5d at 28 DEG C observe the incidence of blade.As shown in Figure 4, Deccan streptomycete QY-3 zymotic fluids pair
Feeling anthracnose rubber tree leaf has preferable control effect, and preventive effect reaches 70%, is significantly higher than the anti-of Bravo (1000 μ g/mL)
Control effect.
The above content is combine specific embodiment further description made for the present invention, it is impossible to assert this hair
Bright specific implementation is confined to these explanations.For those of ordinary skill in the art to which the present invention belongs, do not taking off
On the premise of from present inventive concept, several simple deduction or replace can also be made.
Sequence table
<110>University Of Hainan
<120>One plant of Deccan streptomycete QY-3 and its application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1449
<212> DNA
<213>Deccan streptomycete QY-3 (Streptomyces deccanensis)
<400> 1
gacgaacgct ggcggcgtgc ttaacacatg caagtcgaac gatgaaccac ttcggtgggg 60
attagtggcg aacgggtgag taacacgtgg gcaatctgcc cttcactctg ggacaagccc 120
tggaaacggg gtctaatacc ggatacgaca ctctcgggca tccgatgagt gtggaaagct 180
ccggcggcga aggatgagcc cgcggcctat cagcttgttg gtgaggtaat ggctcaccaa 240
ggcgacgacg ggtagccggc ctgagagggc gaccggccac actgggactg agacacggcc 300
cagactccta cgggaggcag cagtggggaa tattgcacaa tgggcgaaag cctgatgcag 360
cgacgccgcg tgagggatga cggccttcgg gttgtaaacc tctttcagca gggaagaagc 420
gaaagtgacg gtacctgcag aagaagcgcc ggctaactac gtgccagcag ccgcggtaat 480
acgtagggcg cgagcgttgt ccggaattat tgggcgtaaa gagctcgtag gcggtctgtc 540
gcgtcggatg tgaaagcccg gggcttaacc ccgggtctgc attcgatacg ggcagactag 600
agtgtggtag gggagatcgg aattcctggt gtagcggtga aatgcgcaga tatcaggagg 660
aacaccggtg gcgaaggcgg atctctgggc cattactgac gctgaggagc gaaagcgtgg 720
ggagcgaaca ggattagata ccctggtagt ccacgccgta aacggtggga actaggtgtt 780
ggcgacattc cacgtcgtcg gtgccgcagc taacgcatta agttccccgc ctggggagta 840
cggccgcaag gctaaaactc aaaggaattg acgggggccc gcacaagcag cggagcatgt 900
ggcttaattc gacgcaacgc gaagaacctt accaaggctt gacatacacc ggaaacggcc 960
agagatggtc gcccccttgt ggtcggtgta caggtggtgc atggctgtcg tcagctcgtg 1020
tcgtgagatg ttgggttaag tcccgcaacg agcgcaaccc ttgttctgtg ttgccagcat 1080
gcccttcggg gtgatgggga ctcacaggag actgccgggg tcaactcgga ggaaggtggg 1140
gacgacgtca agtcatcatg ccccttatgt cttgggctgc acacgtgcta caatggcagg 1200
tacaatgagc tgcgaagccg tgaggcggag cgaatctcaa aaagcctgtc tcagttcgga 1260
ttggggtctg caactcgacc ccatgaagtc ggagttgcta gtaatcgcag atcagcattg 1320
ctgcggtgaa tacgttcccg ggccttgtac acaccgcccg tcacgtcacg aaagtcggta 1380
acacccgaag ccggtggccc aaccccttgt gggagggagc tgtcgaaggt gggactggcg 1440
attgggacg 1449
Claims (8)
1. one plant of Deccan streptomycete QY-3, which is characterized in that the entitled Streptomyces of Latin of the Deccan streptomycete
Deccanensis, bacterial strain number are QY-3, are preserved in China General Microbiological culture presevation administrative center, deposit number CGMCC
No.14467。
2. Deccan streptomycete QY-3 according to claim 1, which is characterized in that the Deccan streptomycete QY-3 is used to inhibit
Plant pathogenic fungi.
3. Deccan streptomycete QY-3 according to claim 1, which is characterized in that the plant pathogenic fungi includes anthrax-bacilus
Belong to pathogen, banana blight bacteria, fusarium graminearum, cucumber fusarium axysporum, dragon fruit sickle-like bacteria, dragon fruit fester germ,
Botryosphaeria berengeriana f. sp, mango blossom-end rot bacterium or guava intend disk stey.
4. Deccan streptomycete QY-3 according to claim 3, which is characterized in that the colletotrichum pathogen is glue spore charcoal
Subcutaneous ulcer bacterium or Glorosprium musarum Cookeet Mass.
5. a kind of zymotic fluid, which is characterized in that the zymotic fluid prepared by Deccan streptomycete QY-3 described in claim 1 and
.
6. zymotic fluid according to claim 5, which is characterized in that be prepared by the following：To Deccan streptomycete QY-3
Fermented and cultured is carried out, thalline is filtered to remove and obtains zymotic fluid；Fermentation medium is used by fermented and cultured：Millet 20g, albumen
Peptone 3g, glucose sugar 20g, CaCO32g, NaCl 2.5g, water 1000mL, pH7.2-7.4, condition of culture is 28 DEG C, and 180rpm shakes
Swing culture 7d.
7. one plant of Deccan streptomycete QY-3 according to claim 1, which is characterized in that isolated by the following method：
Glacial deposit sample is taken, grinds, is added in the container of built-in sterile water and bead, vibrates, supernatant is taken to carry out
Dilution, obtains dilution, and dilution is taken to be coated on Gao Shi I culture medium flat plates uniformly, cultivates 5-10d, picking actinomyces bacterium colony
It is further isolated and purified using method of scoring, until obtaining actinomyces strain pure culture；Using colletotrichum gloeosporioides Penz as target bacterium, using pair
Stand erect cultivation, screens one plant of bacterial strain QY-3 to colletotrichum gloeosporioides Penz with higher inhibitory activity.
8. one plant of Deccan streptomycete QY-3 according to claim 7, which is characterized in that isolated by the following method：
10g glacial deposit samples are taken, fully grinds, adds in the container of built-in 100mL sterile waters and bead, on shaking table
100rpm vibrates 20min, and supernatant is taken to carry out 10 times of gradient dilutions, obtains dilution, takes dilution 0.1mL in No. I culture of Gao Shi
It is coated with uniformly on base tablet, is placed in 28 DEG C of constant incubators and cultivates 5-10d, picking actinomyces bacterium colony is further using method of scoring
It isolates and purifies, until obtaining actinomyces strain pure culture；Using colletotrichum gloeosporioides Penz as target bacterium, using opposite culture method, screen
One plant of bacterial strain QY-3 to colletotrichum gloeosporioides Penz with higher inhibitory activity.
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| CN110628687A (en) * | 2019-10-24 | 2019-12-31 | 广西科学院 | Streptomyces 5017 and application thereof in antagonism of phytopathogens |
| 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 |
| CN113930355A (en) * | 2021-09-15 | 2022-01-14 | 中国热带农业科学院热带生物技术研究所 | Biocontrol streptomyces weinmannii W7 from termites and application thereof |
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| CN115820454B (en) * | 2022-06-15 | 2023-11-28 | 福建省农业科学院植物保护研究所 | Streptomyces wetland (Streptomyces paludis) strain 13-3 and application thereof |
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