CN101148649B - Bacillus firmus, bacterium agent and application thereof - Google Patents
Bacillus firmus, bacterium agent and application thereof Download PDFInfo
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- CN101148649B CN101148649B CN2007101212746A CN200710121274A CN101148649B CN 101148649 B CN101148649 B CN 101148649B CN 2007101212746 A CN2007101212746 A CN 2007101212746A CN 200710121274 A CN200710121274 A CN 200710121274A CN 101148649 B CN101148649 B CN 101148649B
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Abstract
The present invention discloses one kind of Bacillius firmus SAB-1 preserved in the preservation number of CGMCC No. 2100, and the microbial preparation prepared with Bacillius firmus SAB-1. The Bacillius firmus SAB-1 may be applied in controlling powdery mildew of strawberry, and has the features of high efficiency, broad bacteriostasis spectrum, simple use, low cost, no environmental pollution,etc.
Description
Technical field
The invention belongs to the agricultural microorganism field, relate in particular to a kind of bacillus firmus, and utilize microbiobacterial agent of this bacillus firmus production and its production and application.
Background technology
Powdery mildew of strawberry is the main disease during strawberry produces.In recent years, the generation of powdery mildew of strawberry increases the weight of gradually, particularly protection ground powdery mildew of strawberry; take place when serious, sick leaf rate is more than 45%, and diseased fruit rate is more than 50%; had a strong impact on strawberry output, quality and economic benefit (Yang Lianwei. Yantai fruit tree, 2005 (3): 15～16).Therefore, the peasant claims that powdery mildew of strawberry is " cancer of strawberry ".Powdery mildew of strawberry is caused by the strawberry powdery mildew, the strawberry powdery mildew is garment or robe made of feathers grass single softgel shell (Sphaerotheca aphanis), belongs to Ascomycotina Erysiphales Erysiphaceae list softgel shell and belongs to, and the strawberry Erysiphe is in obligate parasite (Peries O S.Mycologia, 1962,52:380～386).At present, the main method of control powdery mildew of strawberry is to utilize disease-resistant variety and chemical agent.But, the inferior quality of disease-resistant variety generally; Chemical agent control exists again makes germ develop immunity to drugs easily, and produces fruit contamination and problem of environmental pollution, in addition, to the powdery mildew of strawberry prevention effect preferably all azole medicaments all suppress the growth of strawberry.And biological pesticide since selectivity strong, efficient is high, cost is low, free from environmental pollution, to advantages such as person poultry harmless more and more be subjected to people's attention (Liu Bo etc. the Henan agricultural sciences, 2007 (2): 20-23), by screening and utilize the beneficial organism and the meta-bolites thereof of anti-powdery mildew of strawberry, make biological control become the effective way of control powdery mildew of strawberry.
At present; the more effective microorganism or the microbial product of control powdery mildew of strawberry mainly contain: and not streptomyces hygroscopicus Hangzhou mutation (Streptomyces ahygroscopuus v ar.hangzhouensis) (Ye Qiming etc. the Chinese biological control; 2001; 17 (2): envelope 3); the natural compounds osthole (Ze good fortune favour etc. the Laiyang Agricultural College journal; 2005; 22 (3): 189～190 and tight peaceful etc. agricultural chemicals; 2005; 44 (3): 136～137); bacillus cereus TS-02 (Chen Chong etc. Anhui agricultural sciences, 2007,35 (11): 3298～3300).
People have the history in 40 years at least to the research of genus bacillus biological and ecological methods to prevent plant disease, pests, and erosion potential, the field applied research has confirmed the stability of genus bacillus biocontrol fungicide in product, obviously be better than non-genus bacillus and fungi biocontrol microorganisms (.Pest Manag Sci such as Monica L E with the consistency of chemical pesticide and the consistence aspect of different plant different year preventive effects, 2001,57 (8): 695～706.), subtilis (Bacillus subtilis) is the more a kind of antagonistic bacterium of report in the biological control, in the biological control of Plant diseases, bringing into play particularly important effect (He Liyuan. biological control circular .1985,1 (3): 28～31.Kong Jian etc. microorganism journal, 1992,32 (6): 445-449.Lu Suyun. press of Beijing Agricultural University, 1993), it can produce the antibiotic metabolic substd of wide spectrum, it is the superior microorganism population of soil and plant microecology, have very strong anti-adversity ability and antibiotic prophylaxis effect, many good natural strain isolateds successfully are applied to biocontrol of plant disease (Cavaglieri L etc., Researchin Microbiology, 2005.156 (5/6): 748～754), but the effect of relevant bacillus firmus in biological control still rarely reported, (forest-science such as Zhu Tianhui, 2007,43 (2): 120～123). the watchtower sky is quick etc., forestry science and technology, 2007,31 (1): 28～31)) etc. the strain that separation and purification goes out from the soil of loose nursery has the bacillus firmus (B.firmus) of antagonistic activity.Therefore, screening is one of most effectual way of the relevant Plant diseases of control to the inhibited genus bacillus of pathogenic bacteria.Through retrieval, also do not find to be used to prevent and treat the bacillus firmus of powdery mildew of strawberry.
Summary of the invention
One of the object of the invention is the problem that exists in the control powdery mildew of strawberry, and a kind of efficient, active bacillus firmus bacterial strain of broad-spectrum sterilization of having that can be used for the powdery mildew of strawberry biological control is provided.
Second purpose of the present invention provides a kind of microbiobacterial agent that utilizes the production of bacillus firmus bacterial strain and preparation method thereof.
The 3rd purpose of the present invention provided the purposes of bacillus firmus bacterial strain of the present invention on the control powdery mildew of strawberry.
The present invention is achieved through the following technical solutions:
A kind of bacillus firmus SAB-1 (Bacillius firmus SAB-1) is preserved in China Committee for Culture Collection of Microorganisms common micro-organisms center on July 6th, 2007, and deposit number is CGMCC No.2100.
The microbiobacterial agent that utilizes above-mentioned bacillus firmus SAB-1 to produce, its activeconstituents are bacillus firmus SAB-1 thalline and its born of the same parents' extra-metabolite.
The mentioned microorganism microbial inoculum can be liquid formulation.
The preparation method of mentioned microorganism microbial inoculum specifically comprises the steps:
(1) actication of culture: on the SAB-1 inoculation LB plate culture medium with cryopreservation, and under 28～32 ℃, cultivated 20～24 hours; Picking list colony inoculation is on the LB slant medium then, and cultivates 20～24 hours down at 28～32 ℃, washs media surface with sterilized water again, and its elutriant is as inoculation liquid;
(2) preparation of seed liquor: in the LB liquid nutrient medium, insert the inoculation liquid that step (1) prepares according to 0.05～0.5% ratio (volume percent), shake down at 28～34 ℃ and cultivated 12～16 hours, get seed liquor;
(3) fermentation culture: step (2) gained seed liquor is inserted in the Semen Maydis powder soybean cake powder substratum according to 1～10% ratio (volume percent), under 28～34 ℃, shaking speed are the condition of 170～210rpm, cultivated 22～25 hours again, the liquid preparation of bacterial strain SAB-1;
The component and the weight percent thereof of wherein said Semen Maydis powder soybean cake powder substratum are: Semen Maydis powder 2.5%～3.5%, soybean cake powder 2.5%～3.5%, NaCl0.4%～0.6%, CaCO
30.1%～0.2%, KH
2PO
40.01%～0.03% and MgSO
47H
2O 0.02%～0.04%, and all the other are water.
Low temperature described in above-mentioned preparation method's step (1) is meant the used temperature of common preservation bacterial classification, and those skilled in the art according to general knowledge as can be known.
Composition of LB plate culture medium described in above-mentioned preparation method's step (1) or (2), LB slant medium or LB liquid nutrient medium and preparation method thereof all is well known to those skilled in the art, can prepare according to usual method.
The preparation method of above-mentioned Semen Maydis powder soybean cake powder substratum takes by weighing Semen Maydis powder, soybean cake powder, NaCl, CaCO according to weight percent
3, KH
2PO
4And MgSO
4, after their mixing, add water to volume required getting final product again.
Temperature described in above-mentioned preparation method's step (3) is preferably 30～32 ℃, and shaking speed is preferably 210rpm, and incubation time is preferably 24 hours.
The mentioned microorganism microbial inoculum, the viable count of its bacillus firmus SAB-1 is greater than 5 * 10
9Cfu/g.
The application of above-mentioned bacillus firmus SAB-1 on controlling plant diseases.
Above-mentioned bacillus firmus SAB-1 uses on the control powdery mildew of strawberry.
The using method of microbial inoculum of the present invention: with above-mentioned gained microbiobacterial agent dilute with water, liquid bacterial agent after will diluting then carries out blade face, bud and fruit surface spraying before the powdery mildew of strawberry onset peak, also can be sprayed on the face of land on strawberry ground simultaneously, can reach the purpose of control powdery mildew of strawberry.
The screening and separating process of SAB-1 bacterial strain:
The SAB-1 bacterial strain is that Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie separates from the cotton verticillium wilt field soil in Xinjiang and obtains.Soil sample was gathered from 5 in the cotton field on farm, Xinjiang by Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie in 2003; taking by weighing 10g behind the mixing is put in the sterilization triangular flask of 250mL; add the 100mL sterilized water; be put on the shaking table; the 170r/min 30min that vibrates; leave standstill 2h, get supernatant liquor 1mL and add sterilized water 9mL, 10mL10
-2Times soil microorganisms suspension is diluted to 10 with soil supension then
-3, 10
-4, 10
-5, 10
-6Times diluent, each concentration are got 200 μ L microorganism suspensions and are applied on PDA, improvement PDA, LB and the KB culture medium flat plate, and each concentration repeats for 3 times, cultivates 1d, 3d and 5d respectively at 28 ℃ of constant temperature, carries out separation and the purifying of microorganism respectively.And be target control target disease with powdery mildew of strawberry (Sphaerotheca aphanis), Tanaka carries out the screening that the microorganism protection effect is measured in the greenhouse strawberry plantation.The result therefrom filters out a bacterial isolates that the target disease is had remarkable preventive effect, is numbered SAB-1.
The feature of SAB-1 bacterial strain:
(1) morphological specificity
The cell gramstaining of bacterial strain SAB-1 is positive, and thalline is shaft-like, produces gemma behind the cultivation 8h, give birth in the gemma, and ellipse, sporangiocyst does not expand, acid-fast stain feminine gender, no parasporal crystal.On nutrient agar plate, cultivate initial stage bacterium colony light oyster white, the purulence shape, circle, neat in edge, the bacterium colony protuberance is steamed bun shape, surface wettability; Late stage of culture bacterium colony light oyster white, the edge is irregular, and the surface is more moistening, and microscopic folds is arranged; Streak culture on the nutrient agar medium inclined-plane, shape linearly; Static cultivation in the liquid medium within, the surface forms white mycoderm.
(2) 16SrDNA sequencing
The primer of bacterial 16 S rDNA of being used to increase is: F27:(5 ' AGAGTTTGATCATGGCTCAG3 ') with R1492 (5 ' GGCTACCTTGTTACGACTT ' 3).Genomic dna with SAB-1 is template amplification 16S rDNA, and checks order, and the 16SrDNA sequence of SAB-1 is seen sequence table as a result.The 16S rDNA sequence of gained SAB-1 is carried out homology relatively in Genbank, analyze with DNAMAN Version 4.0 according to its 16S rDNA sequence simultaneously and obtained its evolutionary tree (Fig. 1).The homology comparative result shows that the 16S rDNA homology of bacterial strain SAB-1 and subtilis (Bacillus subtilis) and Bacillus licheniformis (Bacillus licheniformis) all reaches 98% (Fig. 2 and Fig. 3), illustrates that the SAB-1 bacterial strain belongs to bacillus (Bacillus).
(3) physiological and biochemical property
The SAB-1 bacterial strain is carried out the Physiology and biochemistry proterties to be detected.Result's (seeing Table 1) shows that the physiological and biochemical property of SAB-1 bacterial strain has the bacillus firmus physiological and biochemical property, illustrates that the SAB-1 bacterial strain belongs to a kind of bacillus firmus.
The conventional identification handbook of bacterium (" general bacterium authentication method commonly used " etc.) show that bacillus firmus is movable, aerobic, Gram-positive, gemma ellipse or column, cyst does not expand, and can grow on the substratum of pH5-7, all can grow in the 2%-10%NaCl substratum; Growth temperature is 10～45 ℃, 28～31 ℃ of optimum growth temperatures.The clark and Lubsreaction feminine gender, the V-P reaction negative.Can not utilize Citrate trianion.The catalase reacting positive, the lecithinase feminine gender.The part Physiology and biochemistry proterties detected result of above SAB-1 bacterial strain is compared with the subtilis reference culture, (Chinese Academy of Sciences's microbial bacterial research division bacteria group is compiled according to " general bacterium authentication method commonly used ", Science Press, 1978), " bacillus ") (Cai Miaoying, war Garrick etc. is translated, agricultural press, 1988) and " microbial taxonomy " (Zhang Jizhong, Fudan University in Shanghai press, 1995) key is retrieved, and determines that the corresponding proterties with bacillus firmus of bacterial strain SAB-1 Physiology and biochemistry proterties is identical.In conjunction with 16S rDNA sequence homology analysis result, identify that finally the SAB-1 bacterial strain is bacillus firmus (Bacillus firmus) simultaneously.
Table 1 bacterial strain SAB-1 Physiology and biochemistry measurement result
Annotate: "+" expression positive findings, "-" expression negative findings, " ND " represents undetermined, " d " expression 11%-89% bacterial strain is positive.
Advantage that the present invention had and beneficial effect: (1) the invention provides a kind of can high-efficiency prevention and control the bacillus firmus bacterial strain of Plant diseases such as powdery mildew of strawberry, a kind of new way of biological control is provided for Plant diseasess such as control powdery mildew of strawberries; (2) biocontrol effect of the present invention is remarkable, and the prevention effect of powdery mildew of strawberry is reached 90.31%～93.42%; (3) preparation method of the present invention is simple, cost is low, be easy to apply; (4) the present invention is pollution-free, and is nuisanceless, and low residue is environmentally friendly, has vital role to improving agricultural product quality and output, can improve the world of agricultural products in China simultaneously and strive power unexpectedly.
Description of drawings
Fig. 1 analyzes the evolutionary tree result who obtains according to the 16SrDNA sequence with DNAMAN 5.2.2.
Fig. 2 is the 16S rDNA gene order homology comparative result of bacillus firmus SAB-1 bacterial strain and subtilis 168 bacterial strains (Bacilliussubtilis 168).Wherein "-" expression is identical with the corresponding base of SAB-1 in B.subtilis 168 and the B.licheniformis ATCC14580 sequence, and " g, a, c, t " represents different with the base in the corresponding site of SAB-1.
Fig. 3 is the 16S rDNA gene order homology comparative result of bacillus firmus SAB-1 bacterial strain and lichem bacillus strain (B.licheniformisATCC 14580).Wherein "-" expression is identical with the corresponding base of SAB-1 in B.subtilis 168 and the B.licheniformis ATCC14580 sequence, and " g, a, c, t " represents different with the base in the corresponding site of SAB-1.
Embodiment
Come further clearly to explain the present invention with specific embodiment below, but be construed as limiting the invention never in any form.Experimental technique among the following embodiment if no special instructions, is ordinary method; Percentage composition among the following embodiment if no special instructions, is weight percentage.
The preparation of SAB-1 microbiobacterial agent, carry out according to following steps:
(1) actication of culture: the bacterial strain SAB-1 that will be stored in-40 ℃ activates (30 ℃) on the LB plate culture medium, and picking list bacterium colony expands numerous (30 ℃) on the LB slant medium cultivates, and gets inoculation liquid; Wherein LB plate culture medium and LB slant medium component and proportioning are: yeast extract 0.5%, and Tryptones 1%, NaCl 0.5%, agar powder 1%; Distilled water 97%, pH7.0-7.4;
(2) preparation of seed liquor: LB nutrient solution 100mL packs in the 250mL triangular flask, the high pressure moist heat sterilization, after treating that temperature drops to room temperature, insert the good SAB-1 inoculation liquid of the above-mentioned activation of step (1) one transfering loop in every bottle, on shaking table, carry out shaking culture, rotating speed 190rpm, and under 30 ℃, cultivated 24 hours, seed liquor got; Wherein LB liquid nutrient medium component and proportioning are: yeast extract 0.5%, and Tryptones 1%, NaCl 0.5%, distilled water 98%, pH7.0-7.4;
(3) preparation of Semen Maydis powder soybean cake powder substratum: according to 3,0% Semen Maydis powder, 3.0% soybean cake powder, 0.5%NaCl, 0.1%CaCO
3, 0.02%KH
2PO
4And 0.03%MgSO
47H
2O prepares substratum, adds water polishing 100% then, mix Semen Maydis powder soybean cake powder substratum; Be sub-packed in the 500mL triangular flask every bottle of 200mL; High pressure moist heat sterilization 30 minutes, cool to again 30 ℃ standby;
(4) fermentation culture: to step (3) gained every bottle of Semen Maydis powder soybean cake powder inoculation of medium step (2) gained seed liquor 2mL; Under 28 ℃ of constant temperature, rotating speed 190rpm condition, carry out fermentation culture;
(5) after the 17h, every 30 minutes from the triangular flask of step (4) sampling carry out microscopy, the gemma in the visual field and total thalline number are counted, and are calculated the gemma rate; The gemma rate reaches 90% can stop fermentation culture; Promptly get bacillus firmus SAB-1 liquid preparation.
Embodiment 2
This test is carried out in the strawberry booth of Baoding, Hebei province Mancheng County peasant household road new website, every sub-district two row, and four repetitions, district's group is arranged at random.Powdery Mildew moderate generation on the strawberry leaves in whole canopy during experiment.(water dilutes, mycetome 10 to adopt knapsack hand sprayer inoculation embodiment 1 prepared bacillus firmus SAB-1 microbial inoculum
8Cfu/mL); Other establishes contrast medicament (10% generation high-moisture shot agent, commercial, Switzerland is just reaching earlier production, 1500 times of water diluent spraying processing) and blank (clear water spraying); Treat the blank " Invest, Then Investigate " of fully falling ill, the powdery mildew of strawberry rank criteria for classifying and preventive effect method of calculation are with reference to " State Standard of the People's Republic of China's pesticide field efficacy medicine test criterion " (two) the 119th parts: bactericidal agent for preventing and treating powdery mildew of strawberry (GB/T17980.119-2004).And calculating disease index and preventive effect.Test-results (seeing Table 2) shows that the prevention effect that the microbial inoculum of bacillus firmus SAB-1 is handled the back powdery mildew of strawberry is significantly higher than blank and medicament contrast, and its preventive effect reaches 93.42%.Bacillus subtilis strain SAB-1 has showed the better prevention effect to powdery mildew of strawberry.
Table 2 bacillus firmus SAB-1 is to the action effect of powdery mildew of strawberry
| Handle | Disease index before handling | Handle the back disease index | Preventive effect (%) |
| SAB-1 | 95.00 | 5.00 | 93.42a |
| Handle | Disease index before handling | Handle the back disease index | Preventive effect (%) |
| Medicament CK | 95.00 | 45.00 | 40.79b |
| Blank CK | 50.00 | 40.00 | 0.00c |
Embodiment 3
This test is carried out at the imperial green science and technology demonstration of Hebei Gaobeidian city sky garden 314 canopies, every sub-district two row, and four repetitions, district's group is arranged at random.Powdery Mildew moderate generation on the strawberry leaves in whole canopy during experiment.(water dilutes, mycetome 10 to adopt knapsack hand sprayer inoculation embodiment 1 prepared bacillus firmus SAB-1 microbial inoculum
8Cfu/mL); Other establishes contrast medicament (10% generation high-moisture shot agent, commercial, Switzerland is just reaching earlier production, 1500 times of water diluent spraying processing) and blank (clear water spraying); Treat the blank " Invest, Then Investigate " of fully falling ill, the powdery mildew of strawberry rank criteria for classifying and preventive effect method of calculation are with reference to " State Standard of the People's Republic of China's pesticide field efficacy medicine test criterion " (two) the 119th parts: bactericidal agent for preventing and treating powdery mildew of strawberry (GB/T17980.119-2004).And calculating sickness rate and preventive effect.Test-results (seeing Table 3) shows that the prevention effect that the microbial inoculum of bacillus firmus SAB-1 is handled the back powdery mildew of strawberry is significantly higher than blank and medicament contrast, and its preventive effect reaches 90.31%.Bacillus subtilis strain SAB-1 has showed the better prevention effect to powdery mildew of strawberry.
Table 3 bacillus firmus SAB-1 is to the action effect of powdery mildew of strawberry
| Handle | Sickness rate (%) before handling | Handle sequela rate (%) | Preventive effect (%) |
| SAB-1 | 70.00 | 5.00 | 90.31a |
| Medicament CK | 75.00 | 35.00 | 36.67b |
| Blank CK | 95.00 | 70.00 | 0.00c |
Embodiment 4
This test is carried out at the imperial green science and technology demonstration of Hebei Gaobeidian city sky garden 308 canopies, every sub-district two row, and four repetitions, district's group is arranged at random.The interior strawberry of whole canopy really goes up the generation of Powdery Mildew moderate during experiment.Processing is divided into thallus suspension liquid and removes thalline born of the same parents extra-metabolite.Bacillus firmus SAB-1 microbial inoculum dilution (mycetome 10 with embodiment 1 gained
8Cfu/mL), then SAB-1 microbial inoculum diluent is divided into two parts, portion is used for direct spraying (being treated to nutrient solution, mycetome and meta-bolites); A under 5000 rev/mins of conditions centrifugal 20 minutes, supernatant liquor was born of the same parents' extra-metabolite of bacillus firmus SAB-1; Precipitation is thallus suspension liquid (mycetome 10 with 0.9% physiological saline suspension of equivalent before centrifugal
8Cfu/mL); With clear water as blank.Result's (seeing Table 4) shows, the prevention effect difference that the nutrient solution of bacillus firmus SAB-1 (mycetome and meta-bolites), born of the same parents' extra-metabolite, thallus suspension liquid are handled is not remarkable, but all be significantly higher than the contrast of blank and medicament, illustrate that bacillus firmus SAB-1 born of the same parents extra-metabolite and viable bacteria body all have preventive and therapeutic effect to powdery mildew of strawberry.Therefore the effective active composition of bacillus firmus SAB-1 control powdery mildew of strawberry comprises thalline and the meta-bolites thereof of bacillus firmus SAB-1.
The outer metabolite of table 4 bacillus firmus SAB-1 thalline and born of the same parents is to the prevention effect of powdery mildew of strawberry
| Handle | Sickness rate (%) before handling | Handle sequela rate (%) | Preventive effect (%) |
| The SAB-1 nutrient solution | 26.79 | 5.00 | 78.37a |
| SAB-1 born of the same parents' extra-metabolite | 20.88 | 8.60 | 52.25a |
| The SAB-1 thallus suspension liquid | 15.10 | 5.00 | 61.63a |
| Medicament CK | 25.90 | 18.33 | 17.98b |
| Blank CK | 20.50 | 17.69 | 0.00c |
Sequence table
＜110〉Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie
＜120〉a kind of bacillus firmus and microbial inoculum thereof and application
<160>1
<170>PatentIn version 3.3
<210>1
<211>1016
<212>DNA
<213>Bacillus firmus
<400>1
ccacttctgt caccttcggc ggctggctcc ataaaggtta cctcaccgac ttcgggtgtt 60
acaaactctc gtggtgtgac gggcggtgtg tacaaggccc gggaacgtat tcaccgcggc 120
atgctgatcc gcgattacta gcgattccag cttcacgcag tcgagttgca gactgcgatc 180
cgaactgaga acagatttgt gggattggct taacctcgcg gtctcgctgc cctttgttct 240
gtccattgta gcacgtgtgt agcccaggtc ataaggggca tgatgatttg acgtcatccc 300
caccttcctc cggtttgtca ccggcagtca ccttagagtg cccaactgaa tgctggcaac 360
taagatcaag ggttgcgctc gttgcgggac ttaacccaac atctcacgac acgagctgac 420
gacaaccatg caccacctgt cactctgccc ccgaagggga agccctatct ctaggggtgt 480
cagaggatgt caagacctgg taaggttctt cgcgttgctt cgaattaaac cacatgctcc 540
accgcttgtg cgggcccccg tcaattcctt tgagtttcag tcttgcgacc gtactcccca 600
ggcggagtgc ttaatgcgtt agctgcagca ctaaggggcg gaaaccccct aacacttagc 660
actcatcgtt tacggcgtgg actaccaggg tatctaatcc tgttcgctcc ccacgctttc 720
gctcctcagc gtcagttaca gaccagagag tcgccttcgc cactggtgtt cctccacatc 780
tctacgcatt tcaccgctac acgtggaatt ccactctcct cttctgcact caagttcccc 840
agtttccaat gaccctcccc ggttgagccg ggggctttca catcagactt aagaaccgcc 900
tgcgagccct ttacgcccaa taattccgga caacgcttgc cacctacgta ttaccgcggc 960
tgctggcacg tagttagccg tgctttctgg ttaggtacgt caaggtgccg ccctat 1016
Claims (9)
1. a bacillus firmus SAB-1 (Bacillius firmus SAB-1) is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, and deposit number is CGMCC No, 2100.
2. the microbiobacterial agent that utilizes the described bacillus firmus SAB-1 of claim 1 to produce is characterized in that its activeconstituents is bacillus firmus SAB-1 thalline and its born of the same parents' extra-metabolite.
3. according to the described microbiobacterial agent of claim 2, it is characterized in that its described microbial inoculum is a liquid formulation.
4. the preparation method of claim 2 or 3 described microbiobacterial agents comprises the steps:
(1) actication of culture: on the SAB-1 inoculation LB plate culture medium with cryopreservation, and under 28～32 ℃, cultivated 20～24 hours; Picking list colony inoculation is on the LB slant medium then, and cultivates 20～24 hours down at 28～32 ℃, washs media surface with sterilized water again, and its elutriant is as inoculation liquid;
(2) preparation of seed liquor: in the LB liquid nutrient medium, insert the inoculation liquid that step (1) prepares according to 0.05～0.5% volume percent ratio, shake down at 28～34 ℃ and cultivated 12～16 hours, get seed liquor;
(3) fermentation culture: step (2) gained seed liquor is inserted in the Semen Maydis powder soybean cake powder substratum according to 1～10% volume percent ratio, cultivated 22～25 hours under 28～34 ℃, shaking speed are the condition of 170～210rpm then, gained is the liquid preparation of bacterial strain SAB-1;
The component and the weight percent thereof of wherein said Semen Maydis powder soybean cake powder substratum are: Semen Maydis powder 2.5%～3.5%, and soybean cake powder 2.5%～3.5%, NaCl 0.4%～0.6%, CaCO
30.1%～0.2%, KH
2PO
40.01%～0.03% and MgSO
47H
2O 0.02%～0.04%, and all the other are water.
5. according to the described preparation method of claim 4, it is characterized in that the described temperature of its step (3) is 30～32 ℃.
6. according to the described preparation method of claim 5, it is characterized in that the described shaking speed of its step (3) is 210rpm.
7. according to the described preparation method of claim 6, it is characterized in that the described incubation time of its step (3) is 24 hours.
8. the application of the described bacillus firmus bacterial strain of claim 1 SAB-1 on controlling plant diseases.
9. the described bacillus firmus bacterial strain of claim 1 SAB-1 uses on the control powdery mildew of strawberry.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106367365A (en) * | 2016-08-24 | 2017-02-01 | 宁波枫叶杰科生物技术有限公司 | Microorganism powder product for treating heavy metal polluted saline-alkali soil and manufacturing method thereof |
| CN106399159A (en) * | 2016-08-30 | 2017-02-15 | 江西顺泉生物科技有限公司 | Bacillus firmus inoculant as well as preparation method and application thereof |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1355292A (en) * | 2000-11-24 | 2002-06-26 | 中国科学院微生物研究所 | Bacillus firmus and its application |
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2007
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1355292A (en) * | 2000-11-24 | 2002-06-26 | 中国科学院微生物研究所 | Bacillus firmus and its application |
Non-Patent Citations (3)
| Title |
|---|
| 刘富平.坚强芽孢杆菌对3种病原真菌的抗生现象.林业科学43 2.2007,43(2),全文. |
| 朱天辉 |
| 朱天辉;刘富平.坚强芽孢杆菌对3种病原真菌的抗生现象.林业科学43 2.2007,43(2),全文. * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106367365A (en) * | 2016-08-24 | 2017-02-01 | 宁波枫叶杰科生物技术有限公司 | Microorganism powder product for treating heavy metal polluted saline-alkali soil and manufacturing method thereof |
| CN106399159A (en) * | 2016-08-30 | 2017-02-15 | 江西顺泉生物科技有限公司 | Bacillus firmus inoculant as well as preparation method and application thereof |
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