CN110093295B - Streptomyces flaviviridis for resisting fish pathogenic bacteria and application thereof - Google Patents
Streptomyces flaviviridis for resisting fish pathogenic bacteria and application thereof Download PDFInfo
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Abstract
The streptomyces flaviviridis for resisting fish pathogenic bacteria and application thereof, the streptomyces flaviviridis is streptomyces flaviviridis X101,Streptomyces flavotricinix101, the strain is preserved in the China center for type culture Collection in 2019, 1 month and 2 days, and the preservation number of the strain is CCTCC NO: m2019004. The invention also comprises the application of the streptomyces flaviviridis, and the streptomyces flaviviridis zymocyte liquid has the bacteriostatic action on fish pathogenic bacteria such as aeromonas veronii, aeromonas hydrophila, aeromonas caviae, erwinia and the like.
Description
Technical Field
The invention relates to a strain of streptomyces flaviviridis (streptomyces flaviviridis) for resisting fish pathogenic bacteriaStreptomyces flavotriciniX101) and application thereof, and biologically determining the screened Streptomyces flaviviridis fermentation broth which has an inhibiting effect on the growth of fish pathogenic bacteria such as Aeromonas veronii, Aeromonas hydrophila, Aeromonas caviae, Erwinia and the like.
Background
Most streptomycetes have antagonistic effects, and over 60 years of research have shown that actinomycetes produce a number of important active natural products that are resistant to cancer, infection and the growth of other harmful microorganisms. Of the approximately 33,500 known bioactive metabolites from bacteria, actinomycetes produce approximately 40%, with over 10,400 from Streptomyces (C.Streptomyces) (see Berdy, J-nos. Thights and efficiencies anti-inflammatory: Where we are now and Where we are doing the same [ J]. Journal of Antibiotics, 2012,65(8):441. ）。
Actinomycetes are by far the most important and most widely used populations of medicinal microorganisms (see Zhangqing, the quarter group, treble, Fanyu, Liuhong soldiers, ZhuTianjiao. secondary metabolites of actinomycetes of marine origin and their biological activities [ J ]. Chinese Marine drugs, 2004, 23(5): 49-54.).
Xantho-tris extractStreptomyces (I), (II)Streptomyces flavotricini，S.f.) is mainly used as antagonistic bacterium of plant pathogenic bacteria, and has relatively obvious effect in antagonizing Myrica rubra canker germ YM 5 strain (see Zheng Jintu, Chengxiaozun, Zhang Xin, Wang Zheng Rong, Xuyongjiang, Hu Tong Hui. antagonistic Myrica rubra canker germ actinomycete screening and identification [ J]Yangzhou university journal (agricultural and life sciences edition), 2015, 36(01).
Streptomyces flaviviridae Y12-26 (Streptomyces flavotriciniS.f. Y12-26) can produce a novel macrolide antifungal antibiotic, bavlomycin K, which has a strong antifungal activity against Pyricularia oryzae (see Zhang DJ, Wei G, Wang Y, Si CC, Tian L, Tao LM, Li YG. Bafilomycin K, a new anti-fingal macrolide from Streptomyces flavotricini Y12-26[J]. Journal of Antibiotics, 2011, 64(5):391-393.)。
At present, streptomyces flaviviridis is used as a fish antagonistic bacterium to inhibit the growth of fish pathogenic bacteria such as aeromonas veronii, aeromonas hydrophila, aeromonas caviae and erwinia, and has not been reported for a while.
Disclosure of Invention
The invention aims to solve the technical problem of overcoming the defects of the prior art and provides a strain of streptomyces flaviviridis for resisting fish pathogenic bacteria and application thereof, so as to effectively prevent and treat fish disease pathogenic bacteria.
The technical scheme adopted by the invention for solving the technical problems is that the streptomyces flavipes X101 (A), (B) and (C) of the inventionStreptomyces flavotriciniX101, abbreviated as s.f.x101), which is cultured in the culture collection center of china (abbreviated as CCTCC, address: wuhan university in Wuhan, China) with the preservation number of the strains being CCTCC NO: m2019004.
The streptomyces flaviviridae 16S rRNA sequence of the invention for resisting fish pathogenic bacteria is as follows: ATGCAGGCGGCTCTACACATGCAGTCGAACGATGAAGCCCTTCGGGGTGGATTAGTGGCGAACGGGTGAGTAACACGTGGGCAATCTGCCCTTCACTCTGGGACAAGCCCTGGAAACGGGGTCTAATACCGGATACGACTGCGGAAGGCATCTTCTGCGGTGGAAAGCTCCGGCGGTGAAGGATGAGCCCGCGGCCTATCAGCTTGTTGGTGGGGTAATGGCCTACCAAGGCGACGACGGGTAGCCGGCCTGAGAGGGCGACCGGCCACACTGGGACTGAGACACGGCCCAGACTCCTACGGGAGGCAGCAGTGGGGAATATTGCACAATGGGCGAAAGCCTGATGCAGCGACGCCGCGTGAGGGATGACGGCCTTCGGGTTGTAAACCTCTTTCAGCAGGGAAGAAGCGAAAGTGACGGTACCTGCAGAAGAAGCGCCGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGGCGCAAGCGTTGTCCGGAATTATTGGGCGTAAAGAGCTCGTAGGCGGCTTGTCACGTCGGATGTGAAAGCCCGAGGCTTAACCTCGGGTCTGCATTCGATACGGGCTAGCTAGAGTGTGGTAGGGGAGATCGGAATTCCTGGTGTAGCGGTGAAATGCGCAGATATCAGGAGGAACACCGGTGGCGAAGGCGGATCTCTGGGCCATTACTGACGCTGAGGAGCGAAAGCGTGGGGAGCGAACAGGATTAGATACCCTGGTAGTCCACGCCGTAAACGTTGGGAACTAGGTGTTGGCGACATTCCACGTCGTCGGTGCCGCAGCTAACGCATTAAGTTCCCCGCCTGGGGAGTACGGCCGCAAGGCTAAAACTCAAAGGAATTGACGGGGGCCCGCACAAGCGGCGGAGCATGTGGCTTAATTCGACGCAACGCGAAGAACCTTACCAAGGCTTGACATATACCGGAAAGCATTAGAGATAGTGCCCCCCTTGTGGTCGGTATACAGGTGGTGCATGGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGCAACCCTTGTCCTGTGTTGCCAGCATGCCCTTCGGGGTGATGGGGACTCACAGGAGACCGCCGGGGTCAACTCGGAGGAAGGTGGGGACGACGTCAAGTCATCATGCCCCTTATGTCTTGGGCTGCACACGTGCTACAATGGCCGGTACAATGAGCTGCGATACCGTGAGGTGGAGCGAATCTCAAAAAGCCGGTCTCAGTTCGGATTGGGGTCTGCAACTCGACCCCATGAAGTCGGAGTCGCTAGTAATCGCAGATCAGCATTGCTGCGGTGAATACGTTCCCGGGCCTTGTACACACCGCCCGTCACGTCACGAAAGTCGGTAACACCCGAAGCCGGTGGCCCAACCCTTGTGGAGGGAGCTTCGAAGTGATGCCTT
Is that
atgcaggcgg ctctacacat gcagtcgaac gatgaagccc ttcggggtgg attagtggcg 60
aacgggtgag taacacgtgg gcaatctgcc cttcactctg ggacaagccc tggaaacggg 120
gtctaatacc ggatacgact gcggaaggca tcttctgcgg tggaaagctc cggcggtgaa 180
ggatgagccc gcggcctatc agcttgttgg tggggtaatg gcctaccaag gcgacgacgg 240
gtagccggcc tgagagggcg accggccaca ctgggactga gacacggccc agactcctac 300
gggaggcagc agtggggaat attgcacaat gggcgaaagc ctgatgcagc gacgccgcgt 360
gagggatgac ggccttcggg ttgtaaacct ctttcagcag ggaagaagcg aaagtgacgg 420
tacctgcaga agaagcgccg gctaactacg tgccagcagc cgcggtaata cgtagggcgc 480
aagcgttgtc cggaattatt gggcgtaaag agctcgtagg cggcttgtca cgtcggatgt 540
gaaagcccga ggcttaacct cgggtctgca ttcgatacgg gctagctaga gtgtggtagg 600
ggagatcgga attcctggtg tagcggtgaa atgcgcagat atcaggagga acaccggtgg 660
cgaaggcgga tctctgggccattactgacg ctgaggagcg aaagcgtggg gagcgaacag 720
gattagatac cctggtagtc cacgccgtaa acgttgggaa ctaggtgttg gcgacattcc 780
acgtcgtcgg tgccgcagct aacgcattaa gttccccgcc tggggagtac ggccgcaagg 840
ctaaaactca aaggaattga cgggggcccg cacaagcggc ggagcatgtg gcttaattcg 900
acgcaacgcg aagaacctta ccaaggcttg acatataccggaaagcatta gagatagtgc 960
cccccttgtg gtcggtatac aggtggtgca tggctgtcgt cagctcgtgt cgtgagatgt 1020
tgggttaagt cccgcaacga gcgcaaccct tgtcctgtgt tgccagcatg cccttcgggg 1080
tgatggggac tcacaggaga ccgccggggt caactcggag gaaggtgggg acgacgtcaa 1140
gtcatcatgc cccttatgtc ttgggctgca cacgtgctac aatggccggt acaatgagct 1200
gcgataccgt gaggtggagc gaatctcaaa aagccggtct cagttcggat tggggtctgc 1260
aactcgaccc catgaagtcg gagtcgctag taatcgcaga tcagcattgc tgcggtgaat 1320
acgttcccgg gccttgtaca caccgcccgt cacgtcacga aagtcggtaa cacccgaagc 1380
cggtggccca acccttgtgg agggagcttc gaagtgatgc ctt 1423
Streptomyces flaviviridae (S.flaviviridus) against fish pathogenic bacteria of the inventionStreptomyces flavotriciniX101, s.f.x 101) identification: directly separating the sludge beside the old vegetable site of the New Zealand institute of Hunan Changsha by adopting a dilution plate coating method to obtain a strain of bacteria with the characteristics of streptomyces flavidus, and identifying the strain as streptomyces flavidus through colony morphology observation, gram staining, physiological and biochemical characteristics and 16S rRNA gene homology analysisStreptomycesBelongs to the field of chemical engineering and belongs to the technical field of chemical engineering,flavotricinithe strain is named as streptomyces flavacinus X101 (S.f. X101).
Antibacterial activity:
inoculating the streptomyces flaviviridus X101 into a fermentation culture medium, and performing fermentation culture at the temperature of 30-32 ℃ and the rotational speed of 180-. Taking zymocyte liquid, and performing antibacterial activity experiments on Aeromonas veronii, Aeromonas hydrophila, Aeromonas caviae and Erwinia.
The fermentation medium comprises the following components: 20 g/L of starch, 10 g/L of glucose, 5 g/L of bacteriological peptone, 5 g/L of yeast extract, 5 g/L of calcium carbonate, 7.2 +/-0.2 of pH and 20min of sterilization at 115 ℃.
The streptomyces flaviviridis is taken as an important plant pathogen antagonistic bacterium, the action range of the streptomyces flaviviridis taken to cover pathogenic bacteria of rice blast and red bayberry canker, the streptomyces flaviviridis obtained by separation of the streptomyces flaviviridis has good bacteriostatic effect on pathogenic bacteria of fishes such as aeromonas veronii, aeromonas hydrophila, aeromonas caviae, erwinia and the like, and the streptomyces flaviviridis has important application value on disease control of cultured fishes.
Streptomyces flaviviridis X101 is obtained by screening in the laboratory for the first time, and the invention reports that Streptomyces flaviviridis is taken as fish antagonistic bacteria to inhibit the growth of fish pathogenic bacteria such as aeromonas veronii, aeromonas hydrophila, aeromonas caviae and erwinia.
Drawings
FIG. 1 is a gram staining morphological feature diagram of the strain S.f.X101 after separation and purification;
FIG. 2 is a scanning electron microscope morphological characteristic diagram of the strain S.f.X101 after separation and purification;
fig. 3 is a tree of s.f. x101 strain 16S rRNA sequences evolved;
FIG. 4 is a graph showing the growth of S.f.X101 strain in Streptomyces sp culture medium No. 1 (abbreviation: ISP 1);
FIG. 5 is a graph of bacteriostatic results of fermentation supernatants after 120h of S.f.X101 culture;
FIG. 6 is an observation picture of pathological phenomena in vivo detection of Streptomyces flavotricins activity against fish pathogenic bacteria; (CK is healthy grass carp; A.v is grass carp infected by only using aeromonas veronii injection, the body surface, anus, fin base and abdomen of the grass carp have obvious hyperemia symptom, and the red and swollen anus is serious; A.v + S.f.X101 is grass carp injected with aeromonas veronii after being soaked by streptomyces xanthus fermentation liquor, and the hyperemia symptom of the grass carp is relieved);
FIG. 7 is a comparison and observation chart of H & E staining of renal tissue in vivo detection of Streptomyces flaviviridae anti-fish pathogenic bacteria activity; (CK is a healthy grass carp kidney sample; A.v is a grass carp kidney sample infected by using only the aeromonas veronii injection; A.v + S.f. X101 is a grass carp kidney sample injected with the aeromonas veronii after being soaked and treated by the streptomyces xanthus fermentation liquor);
FIG. 8 is a graph showing H & E staining comparison of liver tissue in vivo detection of Streptomyces flavotricin activity against fish pathogenic bacteria; (CK is a healthy grass carp liver sample; A.v is a grass carp liver sample infected by using only the aeromonas veronii injection; A.v + S.f. X101 is a grass carp liver sample injected with the aeromonas veronii after being soaked and treated by the streptomyces xanthus fermentation liquor).
Description of the preservation of the microorganism strains
Streptomyces flaviviridae X101 (S. flaviviridus)Streptomyces flavotriciniX101), the strain is preserved in the China center for type culture Collection in 2019, 1 month and 2 days, and the preservation number of the strain is CCTCC NO: m2019004.
Detailed Description
The present invention will be described in further detail with reference to specific examples.
Examples
The streptomyces flaviviridis X101 (S.f. X101) of the embodiment is preserved in a China center for type culture Collection (CCTCC for short, address: Wuhan university in Wuhan, China) in 2019, 1 month and 2 days, and the preservation number of the strain is CCTCC NO: m2019004.
The separation process of the streptomyces flavidus X101 strain of the embodiment: collecting soil samples from the old vegetable site of the New society of Hunan province, adding sterile water, mixing uniformly, diluting in a gradient manner, uniformly coating 100 microliters (mu L) of the soil samples on a culture medium plate No. 1 Gauss, selecting a single colony from the plate, diluting again and scribing on the culture medium plate No. 1 Gauss, culturing for 72h at the temperature of 30 ℃, inoculating the single colony which is repeatedly separated and purified into a shake flask of a liquid culture medium No. 1 Gauss, culturing for 72h at the speed of 30 ℃, 180-plus-200 rpm, mixing 50% glycerol uniformly, and preserving for a long time in a refrigerator at the temperature of-80 ℃.
Observation by an optical microscope and an electron microscope:
the specific implementation process comprises the following steps: taking 1.5mL of S.f.X101 bacterial solution cultured for 72h, 9000rpm for 3 min, centrifuging, removing supernatant, washing twice with double distilled water, re-suspending the bacteria with 200 mu L of double distilled water, sucking 10 mu L of bacterial solution, dripping the bacterial solution in the center of a glass slide, fixing a smear by flame for 1-2 times, adding crystal violet, dyeing for 1min, and washing with water; adding 95% alcohol, shaking the slide, decolorizing for 20-60s according to the thickness of the slide, washing with water, and sucking off water; adding safranin, dyeing for 1min, and washing with water; after the water was blotted dry, the oil was examined under a microscope. The morphology of the cells was observed under 100-fold oil-scope. Taking S.f. X101 to culture 72h of bacterial liquid 1.5mL, 9000rpm for 3 min, centrifuging, removing supernatant, washing with physiological saline for 3 times, washing with PBS buffer for 6-8 times, finally resuspending the bacterial cells with 200 mu L of PBS buffer, adding 2.5% glutaraldehyde fixing liquid into washed precipitates for 12h, washing with sterile ultrapure water for 2 times, then sequentially dehydrating with 30%, 50%, 70%, 80% and 90% ethanol in mass concentration gradients, standing for 15 min in each gradient, dehydrating with absolute ethanol for 2 times, standing for 15 min each time after finishing dehydration, finally sucking the uniformly mixed sample-ethanol suspension liquid to drip on a sample platform covered with a cover glass, and observing the shape of the bacterial cells under a scanning electron microscope.
Gram-stained microscopic pictures of strain s.f.x101 cultured for 72h (see fig. 1), showing that the strain is gram-positive. X101 (see fig. 2).
Analysis of homologous sequences of 16s rRNA genes of S.f. X101 strain:
the specific implementation process comprises the following steps: a single colony is picked from a Gauss No. 1 culture medium plate, transferred into a 1.5mL Ep tube containing 1.2 mL of a Gauss No. 1 liquid culture medium, a small hole is punctured on a tube cover by using a syringe needle, the temperature is 30 ℃, the rpm is 850, after shaking culture is carried out for 72h, a bacterial genome DNA extraction kit is used, and whole genome extraction is carried out according to the operation steps. General primers (Bf-F, AGAGTTTGATCCTGGCTCAG; Bf-R, ACGGCTACCTTGTTACGACTT) were designed based on the bacterial 16S rRNA gene sequence and synthesized by Biotechnology Inc., Shanghai.
16S rRNA gene amplification was performed according to the following reaction system and reaction conditions:
reaction system (20 μ L): sterile double distilled water, 12 μ L; 5 × Buffer, 4 μ L; dNTP, 1.6. mu.L; Bf-R (10 mu M), 0.6 mu L; Bf-F (10. mu.M), 0.6. mu.L; genome template, 1 μ L; PrimerSTAR DNA Polymerase (Takara), 0.2. mu.L;
reaction procedure: pre-denaturation at 94 deg.C for 4 min, denaturation at 94 deg.C for 30 s, annealing at 52 deg.C for 30 s, extension at 72 deg.C for 90 s, 30 cycles, and extension at 72 deg.C for 10 min.
The PCR product is purified by a multifunctional DNA purification recovery kit and sent to the handed-in (Shanghai) biotechnology limited company for sequencing together with a proper amount of primers. The sequencing result showed that the 16S rRNA gene sequence of the isolated strain S.f.X101 was 1423 bp in length. The 16S rRNA gene sequences of S.f.X101 were aligned by on-line BLAST (NCBI, http:// www.ncbi.nlm.nih.gov), and homology analysis of the 16S rRNA gene sequences revealed that the strains were respectively compared with those of the S.f.X101Streptomyces flavotricini NYMH12 、Streptomyces amritsarensis BTU8、Streptomyces polychromogenes DC19、Streptomyces toxytricini HBC11、Streptomyces polychromogenes The similarities of DC-19 and the like are all 99%. The strain was presumed to belong to based on the BLAST results and phase contrast microscopyStreptomyces flavotriciniSubspecies, named asStreptomyces flavotriciniX101 and a phylogenetic tree was constructed (see FIG. 3).
⑤Streptomyces flavotriciniGrowth curve determination for X101:
gao's No. 1 medium: soluble starch 20 g/L, KNO 3 1 g/ L，K 2 HPO 4 0.5 g/ L，MgSO4·7H 2 O 0.5 g/ L，NaCl 0.5 g/ L，FeSO 4 · 7H 2 O 0.01 g/ L，pH=7.4-7.6；
The streptomyces culture medium No. 1 comprises the following components: casein peptone 5 g/L, yeast extract 3 g/L, pH =7.0 ± 0.2.
The specific implementation process comprises the following steps: picking from Gao's No. 1 flat plateTaking a single colony, transferring the single colony into a 1.5mL Ep tube containing 1.2 mL of the Gao's No. 1 liquid culture medium, pricking a small hole on a tube cover by using a syringe needle, performing shaking culture at 30 ℃, 850 rpm and 72 hours, transferring the single colony into a 30mL streptomyces culture medium No. 1 at 30 ℃,180 and 200rpm, performing shaking culture every 12 hours, and measuring OD (optical density) 600 The value is obtained.
The growth curve result of the streptomyces flaviviridis X101 strain in the fermentation medium (see figure 4) shows that the lag phase of the same strain is 24 hours initially, the logarithmic phase is 24-72 hours, the 72-108 hours are stationary phases, and the stationary phase of the strain is short. The bacteriostatic effect is best in the stationary phase, and the number of thalli is close to the maximum. The nutrient substances are consumed to be exhausted, which is not beneficial to the growth of the bacteria. After 108 h, the cell is in a decay period, the number of viable bacteria in the decay period is reduced, the bacteria begin to dissolve, and the physiological metabolic activity basically stops.
Sixthly, determining the bacteriostatic activity of the streptomyces flavotricin X101 strain:
the specific implementation process comprises the following steps: after the streptomyces flaviviridis X101 strain is activated in a Gao's No. 1 culture medium for 72h, the strain is respectively inoculated into a fermentation culture medium by 1 percent of inoculation amount, the culture is carried out for 120h, and fermentation liquor is collected. 100 mu L of the bacterial liquid of Aeromonas veronii, Aeromonas hydrophila, Aeromonas caviae and Erwinia are taken and coated on an LB agar plate, 10 mu L of the centrifuged supernatant of the fermentation liquid is dripped on a filter paper sheet, the diameter of the filter paper sheet is 6 mm, the filter paper sheet is cultured for 8-12 h at 30 ℃, and the antibacterial activity is measured (see figure 5).
The result shows that the streptomyces flaviviridis X101 fermentation supernatant has good bacteriostatic effect on fish pathogenic bacteria such as Aeromonas veronii, Aeromonas hydrophila, Aeromonas caviae, Erwinia and the like.
The fermentation medium comprises the following components: 20 g/L of starch, 10 g/L of glucose, 5 g/L of bacteriological peptone, 5 g/L of yeast extract, 5 g/L of calcium carbonate and pH 7.2 +/-0.2.
Seventhly, detecting the activity of the streptomyces flavotrivinum on fish pathogenic bacteria in vivo:
the specific implementation process comprises the following steps: selecting grass carps of the same strain and with consistent size from a Tanchet-Taoist fish breeding base, wherein the size is 12-15 cm; the body weight is in the range of 35g-55 g. The water used was aerated at least before the experiment72h, feeding the grass carps normally for one to two weeks, and randomly dividing the grass carps into 3 groups; the treatment group treats the water body by using the streptomyces flavipes fermentation supernatant with the concentration higher than the minimum bacteriostatic concentration, controls the final concentration of the fermentation liquor in the water body to be 800-1000 mu g/mL, and puts the grass carps into the water body added with the streptomyces flavipes fermentation supernatant for soaking treatment for 12 hours; and (3) placing the control group into a water body added with the fermentation culture medium with the same concentration for treatment for 12 hours. Using 200 μ L LD 50 A.v the two groups of fishes injected with Aeromonas veronii were observed and compared with the third group of healthy grass carps without treatment (see fig. 6), and the pathological symptoms of congestion of body surface, anus, fin base and abdomen were found to be relieved after the treated group of grass carps injected with Aeromonas veronii compared with the control group. Liver and kidney tissue samples of three groups of fishes were taken at 24H, fixed with 4% paraformaldehyde, dehydrated and embedded in paraffin, and sections (5 μm) were prepared for H & E staining observation (see FIG. 7; FIG. 8), and it was found that the glomeruli and renal tubules of the kidney tissue of the control group were disintegrated, whereas the pathological symptoms of the grass carp group treated with the Streptomyces flaviperidans fermentation supernatant in advance were alleviated, consistent with the kidney of normal fish bodies, and no significant disintegration occurred. Also, the disintegration of liver lobules in the treated group was also alleviated compared to the control group in liver tissue.
Sequence listing
<110> university of Hunan province
<120> Streptomyces flaviviridis for resisting fish pathogenic bacteria and application thereof
<130>
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<211> 1423
<212> DNA/RNA
<213>Streptomyces flaviviridae X101Streptomyces flavotricini X101
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atgcaggcgg ctctacacat gcagtcgaac gatgaagccc ttcggggtgg attagtggcg 60
aacgggtgag taacacgtgg gcaatctgcc cttcactctg ggacaagccc tggaaacggg 120
gtctaatacc ggatacgact gcggaaggca tcttctgcgg tggaaagctc cggcggtgaa 180
ggatgagccc gcggcctatc agcttgttgg tggggtaatg gcctaccaag gcgacgacgg 240
gtagccggcc tgagagggcg accggccaca ctgggactga gacacggccc agactcctac 300
gggaggcagc agtggggaat attgcacaat gggcgaaagc ctgatgcagc gacgccgcgt 360
gagggatgac ggccttcggg ttgtaaacct ctttcagcag ggaagaagcg aaagtgacgg 420
tacctgcaga agaagcgccg gctaactacg tgccagcagc cgcggtaata cgtagggcgc 480
aagcgttgtc cggaattatt gggcgtaaag agctcgtagg cggcttgtca cgtcggatgt 540
gaaagcccga ggcttaacct cgggtctgca ttcgatacgg gctagctaga gtgtggtagg 600
ggagatcgga attcctggtg tagcggtgaa atgcgcagat atcaggagga acaccggtgg 660
cgaaggcgga tctctgggccattactgacg ctgaggagcg aaagcgtggg gagcgaacag 720
gattagatac cctggtagtc cacgccgtaa acgttgggaa ctaggtgttg gcgacattcc 780
acgtcgtcgg tgccgcagct aacgcattaa gttccccgcc tggggagtac ggccgcaagg 840
ctaaaactca aaggaattga cgggggcccg cacaagcggc ggagcatgtg gcttaattcg 900
acgcaacgcg aagaacctta ccaaggcttg acatataccggaaagcatta gagatagtgc 960
cccccttgtg gtcggtatac aggtggtgca tggctgtcgt cagctcgtgt cgtgagatgt 1020
tgggttaagt cccgcaacga gcgcaaccct tgtcctgtgt tgccagcatg cccttcgggg 1080
tgatggggac tcacaggaga ccgccggggt caactcggag gaaggtgggg acgacgtcaa 1140
gtcatcatgc cccttatgtc ttgggctgca cacgtgctac aatggccggt acaatgagct 1200
gcgataccgt gaggtggagc gaatctcaaa aagccggtct cagttcggat tggggtctgc 1260
aactcgaccc catgaagtcg gagtcgctag taatcgcaga tcagcattgc tgcggtgaat 1320
acgttcccgg gccttgtaca caccgcccgt cacgtcacga aagtcggtaa cacccgaagc 1380
cggtggccca acccttgtgg agggagcttc gaagtgatgc ctt 1423
Claims (5)
1. The streptomyces flaviviridis for resisting fish pathogenic bacteria, and is characterized in that the streptomyces flaviviridis is streptomyces flaviviridis X101 (Streptomyces flaviviridis X101)Streptomyces flavotriciniX101), the strain is preserved in the China center for type culture Collection in 2019, 1 month and 2 days, and the preservation number of the strain is CCTCC NO: m2019004.
2. The fish pathogen-resistant Streptomyces flaviviridae according to claim 1, wherein the 16S rRNA sequence thereof is represented by SEQ ID NO. 1 of the sequence Listing.
3. Use of the fish pathogenic bacterium-resistant Streptomyces flaviviridae according to claim 1 or 2 for the manufacture of a medicament for inhibiting the fish pathogenic bacteria Aeromonas veronii, Aeromonas hydrophila, Aeromonas caviae, Erwinia.
4. The use of Streptomyces flaviviridae against fish pathogenic bacteria according to claim 3, wherein the bacteriostatic agent is a fermented bacterial liquid, and the preparation method comprises: inoculating the streptomyces flaviviridus X101 into a fermentation culture medium, and performing fermentation culture at the temperature of 30-32 ℃ and the rotational speed of 180-.
5. The use of Streptomyces flaviviridae against fish pathogens according to claim 4 wherein the fermentation medium comprises the following components: 20 g/L of starch, 10 g/L of glucose, 5 g/L of bacteriological peptone, 5 g/L of yeast extract, 5 g/L of calcium carbonate, 7.2 +/-0.2 of pH and 20min of sterilization at 115 ℃.
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|---|---|---|---|---|
| JPS55118490A (en) * | 1979-03-07 | 1980-09-11 | Takeda Chem Ind Ltd | Demethylmaytansinoid compound and its preparation |
| CN103205389A (en) * | 2013-05-13 | 2013-07-17 | 扬州大学 | Streptomyces flavotricini for antagonizing botryosphaeria dothidea |
| CN107058178A (en) * | 2017-03-13 | 2017-08-18 | 陕西博秦生物工程有限公司 | A kind of plain streptomycete of Huang three and microbial inoculum and its application |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS55118490A (en) * | 1979-03-07 | 1980-09-11 | Takeda Chem Ind Ltd | Demethylmaytansinoid compound and its preparation |
| CN103205389A (en) * | 2013-05-13 | 2013-07-17 | 扬州大学 | Streptomyces flavotricini for antagonizing botryosphaeria dothidea |
| CN107058178A (en) * | 2017-03-13 | 2017-08-18 | 陕西博秦生物工程有限公司 | A kind of plain streptomycete of Huang three and microbial inoculum and its application |
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