CN109207393A - One bacillus amyloliquefaciens and its application in prevention and treatment celery root rot - Google Patents

One bacillus amyloliquefaciens and its application in prevention and treatment celery root rot Download PDF

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CN109207393A
CN109207393A CN201811052395.4A CN201811052395A CN109207393A CN 109207393 A CN109207393 A CN 109207393A CN 201811052395 A CN201811052395 A CN 201811052395A CN 109207393 A CN109207393 A CN 109207393A
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bacillus amyloliquefaciens
root rot
strain
prevention
control agent
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牛兆颖
王凤娇
张淑凤
刘新
李宁
邱振豪
张淑静
牛赡光
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Qingdao Nuubel Biological Engineering Co ltd
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Abstract

The invention discloses a bacillus amyloliquefaciens and its applications in prevention and treatment celery root rot.The bacterial strain is bacillus amyloliquefaciens (Bacillus amyloliquefaciens) TJ1, and deposit number is CGMCC No.14741.Experiment shows that Bacillus amyloliquefaciens strain TJ1 of the invention has that the speed of growth is fast, sporulation quantity is big, resistance is strong, it is good to prevent and treat celery root rot effect, Celery rhizosphere can rapid, high volume colonize the features such as, therefore have a good application prospect.The biological control agent prepared by Bacillus amyloliquefaciens strain TJ1, can be used as biological pesticide or bio-feritlizer, and prevention and treatment celery root rot control efficiency is up to 80% or more.

Description

One bacillus amyloliquefaciens and its application in prevention and treatment celery root rot
Technical field
The invention belongs to biological control plant disease fields.Specifically, the present invention relates to a bacillus amyloliquefaciens And its application in prevention and treatment celery root rot, and the biological control agent using bacillus amyloliquefaciens preparation.
Background technique
Celery root rot is mainly the plant disease as caused by Fusarinm solani, is a kind of soil-borne disease, main harm celery The basal part of stem of dish plant and root.After celery is infected, occurred the wine-colored scab of water soaking mode first by infringement position, it is several Scab color gradually becomes dark brown or dark brown after it.Subsequent scab starts to be recessed, and blade becomes yellow, and illness is from lower to upper Gradually develop.In addition, celery root rot frequently results in celery plant main root decay and necrosis, lateral root growth is influenced, last celery is whole Strain is wilted withered.
Celery root rot is resulted in significant economic losses to peasant household.Prevention and treatment for celery root rot, the side routinely used Method includes traditional cultural control, chemical prevention, cultivates disease-resistant variety and biological control.Cultural control such as efficent rotation is kept away Exempt from mechanical damage, reinforce cultivation management while to deep plough, reduces and prevent the gas permeability for increasing soil using single nitrogenous fertilizer, but Heavy workload, investment are high.Chemical prevention can use 50% carbendazol wettable powder, 500 times of liquid or 70% when disease just occurs Fenaminosulf wettable powder etc. carries out pouring root treatment.This method efficacy time is short, usage amount is big, easily causes pesticide residue, ring Border pollution；Meanwhile the long-time service of chemical pesticide leads to preventive effect decline even failure so that pathogen develops drug resistance to it. The disease-resistant variety R&D cycle is long, and investment is big, promotes slow.Biological control mainly utilizes beneficial microorganism, by interbiotic Competition, antibacterial action, hyperparasitism, cross-protection and induced resistance etc. inhibit the survivals of certain pathogens And activity, have the advantages that it is nontoxic, pollution-free, do not develop drug resistance, be efficient, show wide application prospect.
Bacillus is a kind of saprophytic bacteria being distributed widely in various different living environments, is the aerobic simultaneous anaerobism of a group Production gemma Gram-positive bacillus general name, can produce the inverse gemma of interior raw heat-resistant.Gemma not only has resistance to heat, Also there is very strong resistance to ultraviolet light, electromagnetic radiation and some chemicals, it can endure various poor environments.Due to tool Have so strong resistance so that it is conducive to production and the formulation of biocontrol agent, be also conducive to its survival in the environment, It colonizes and breeds.Since they grow, fast, nutrition is simple, is capable of forming the gemma with stronger resistance ability, opens in product The advantages such as more non-bacillus living bacteria count amount is high in hair, performance is stablized and attract attention.The solution starch of bacillus subordinate Bacillus is with the following functions in agricultural production: 1, disease-resistant antibacterial broad-spectrum high efficacy: having significant preventive effect to soil-borne disease；2, Degeneration-resistant health care promotes growth: improving crop anti-adversity, promotes crop Rapid Rooting, improve root hydraulic conductivity, promotes plant strong Kang Shengchang；3, improvement soil promotes fertility: improve crop rhizosphere Tiny ecosystem, the potential nutrients such as phosphorus, potassium of indissoluble in activating soil, Soil is improved, it is loose hardened, contain soil degradation, increases soil fertility, the use for improving and extending fertilizer efficiency, reduce chemical fertilizer Amount；4, reduce the residual high-quality volume increase of agriculture: the pesticide residue in degradation soil and fruit reduces The sub-salt content, improves fruits and vegetables vitamin And sugared content, improve quality of agricultural product, improves crop yield, easily storage transport.
Summary of the invention
The object of the present invention is to provide one plant for efficiently preventing and treating the new bacillus amyloliquefaciens of celery root rot --- Bacillus amyloliquefaciens strain TJ1.Bacterial strain of the invention is with the speed of growth is fast, sporulation quantity is big, control efficiency is good, resistance By force, plant rhizosphere can rapid, high volume colonize the features such as, therefore have a good application prospect.
Bacterial strain provided by the present invention is bacillus amyloliquefaciens (Bacillus amyloliquefaciens) TJ1, is Separation obtains from Shandong Province's Jiaodong Peninsula compost sample, has been preserved in Chinese microorganism strain guarantor on September 26th, 2017 Hide administration committee's common micro-organisms center (abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese section Institute of microbiology of institute), deposit number is CGMCC No.14741.
The cultural method or numerous of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) bacterial strain TJ1 of the present invention Growing method includes:
(1) Nostoc commune Vanch, which saves, uses LB culture medium, formula are as follows: tryptone 10g, yeast extract 5g, sodium chloride 10g, agar 15-20g, distilled water 1000mL, pH are adjusted to 7.0-7.2；
(2) laboratory fluids culture uses LB liquid medium, formula are as follows: tryptone 10g, yeast extract 5g, chlorine Change sodium 10g, distilled water 1000mL, pH are adjusted to 7.0-7.2；
(3) seed liquor culture medium prescription: by mass percentage, beancake powder 3.00%, corn flour 4.00%, glucose 0.40%, fish meal 0.50%, potassium dihydrogen phosphate 0.03%, precipitated calcium carbonate 1.20%, ammonium sulfate 0.30%, sodium chloride 0.10%, remaining as water, pH is adjusted to 7.0-7.2.
(4) bulk fermentation culture medium prescription: with seed liquor culture medium prescription in (3).
The invention also discloses a kind of biological control agents including the Bacillus amyloliquefaciens strain TJ1.
The preparation method of biological control agent of the present invention the following steps are included:
(1) seed liquor of Bacillus amyloliquefaciens strain TJ1 is prepared；
(2) seed liquor prepared by step (1) is inoculated into an aseptic environment in sterilized bulk fermentation culture medium, Constant temperature incubation 28h-32h at 26-30 DEG C；
(3) carrier will be added in the fermentation liquid of step (2) culture, spray drying obtains bacillus amyloliquefaciens TJ1 bacterium powder；
(4) agriculturally common auxiliary agent such as wetting agent, filler can be further added in bacillus amyloliquefaciens TJ1 bacterium powder Etc. biological control agent is made.
The present invention also provides above-mentioned Bacillus amyloliquefaciens strain TJ1 or above-mentioned biological control agent in prevention and treatment celery Purposes in root rot.
Experiment shows that Bacillus amyloliquefaciens strain TJ1 of the invention is fast with the speed of growth, sporulation quantity is big, resistance By force, Celery rhizosphere can rapid, high volume colonize the features such as, therefore have a good application prospect.By the bacillus amyloliquefaciens The biological control agent of bacterial strain preparation, can be used as biological pesticide or bio-feritlizer, efficiently prevents and treats celery root rot, is a kind of pole Has the biocontrol agent of application prospect.
Specific embodiment
The invention will now be further described with reference to specific embodiments, the advantages and features of the present invention will be with description and It is apparent.But these embodiments are merely exemplary, and it is not intended to limit the scope of the present invention in any way.Those skilled in the art Member it should be understood that without departing from the spirit and scope of the invention can details to technical solution of the present invention and form into Row modifications or substitutions, but these modifications and replacement are fallen within the protection scope of the present invention.
Embodiment 1
1, the isolation and purification of Bacillus amyloliquefaciens strain TJ1
Bacillus amyloliquefaciens strain TJ1 of the invention is to use dilution-plate method from Shandong Province's Jiaodong Peninsula compost sample Acquisition, separation method are separated with plate streak are as follows:
(1) the separation of bacillus: the acquisition of sample collects 200 parts of compost samples in Jiaodong Peninsula in Shandong Province.Claim It takes 1g sample in 100mL sterile water, is placed in 150rpm in 30 DEG C of shaking tables and shakes 10min, be subsequently placed in 60 DEG C of water-baths and incubate 30min is educated, 100 μ L 10 are taken-2、10-3、10-4Dilution is coated on LB culture medium flat plate, and each gradient is coated with three in parallel, It falls in the microbial bacteria that 30 DEG C are cultivated the different shape after 2d on picking LB culture medium and crosses on LB culture medium flat plate, it is fixed When observe bacterium colony growing state.Then plate streak is used, Bacillus strain is purified, number saves respectively.
(2) the screening of the efficient antagonistic strain of celery root rot
1. primary dcreening operation: using opposite culture method, prepare LB plate, beaten with punch at bacillus, celery root rot edge Taking diameter is the bacteria cake of 5mm, is implanted in the opposite two sides center of plate respectively, and 25 DEG C of constant temperature incubations observe bacillus day by day To the inhibiting effect of pathogen.
2. secondary screening: the Bacillus strain with efficient antagonistic activity screened being carried out secondary screening, mainly by resistance to Warm nature, resistance to acid and alkali, drug-resistant test screen the preferable Bacillus strain of patience, carry out potting controlling experiment.
The present inventor obtains one plant of solution starch gemma bar that can efficiently prevent and treat celery root rot by a large amount of screening operations Bacteria strain TJ1.It is demonstrated experimentally that the bacillus amyloliquefaciens original powder shows very efficient prevention and treatment effect in prevention and treatment celery root rot Fruit increases production it significantly.Thus, bacillus amyloliquefaciens of the invention are the bacillus amyloliquefaciens with wide application prospect New strains can be used for preparing the biological control agent of prevention and treatment celery root rot.
2, bacterial strain is identified
(1) Microbiological Characteristics: colonial morphology is circle in beef extract-peptone agar medium or LB culture medium, flat It is flat, toughness is moistened, White-opalescent, rough surface edge is irregular, in beef extract-peptone agar medium, at 28 DEG C Two days later, microscopy, somatic cells are in rod-short for culture, can be moved.It is positive through Gram's staining (with Escherichia coli for control). Endogenous spore can be formed, gemma expands, and it is oval that gemma is observed after spore staining.
(2) molecular biological characteristic
The 16s rDNA result of gene sequence determination of the bacterial strain is following (SEQ-1):
CTAGAGGTTACCTTACCGACTTCGGGTGTTACAAACTCTCGTGGTGTGACGGGCGGTGTGTACAAGGC CCGGGAACGTATTCACCGCGGCATGCTGATCCGCGATTACTAGCGATTCCAGCTTCACGCAGTCGAGTTGCAGACT GCGATCCGAACTGAGAACAGATTTGTGGGATTGGCTTAACCTCGCGGTTTCGCTGCCCTTTGTTCTGTCCATTGTA GCACGTGTGTAGCCCAAGGTATAAGGGGGCATGATGATTTGACGTCATCCCCACCTTCCTCCGGTTTGTCACCGGC AGTCACCTTAGAGTGCCCAACTGAATGCTGGCAACTAAGATCAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACA TCTCACGACACGAGCTGACGACAACCATGCACCACCTGTCACTCTGCCCCCGAAGGGGACGTCCTATCTCTAGGAT TGTCAGAGGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGG GCCCCCGTCAATTCCTTTGAGTTTCAGTCTTGCGACCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGC ACTAAGGGGCGGAAACCCCCTAACACTTAGCACTCATCGTTTACGGCGTGGACTACCAGGGTATCTAATCCTGTTC GCTCCCCACGCTTTCGCTCCTCAGCGTCAGTTACAGACCAGAGAGTCGCTTTCGCCACTGGTGTTCCTCCACATCT CTACGCATTTCACCGCTACACGTGGAATTCCACTCTCCTCTTCTGCACTCAAGTTCCCCAGTTTCCAATGACCCTC CCCGGTTGAGCCGGGGGCTTTCACATCAGACTTAAGAAACCGCCTGCGAGCCCTTTACGCCCAATAATTCCGGACA ACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGGCTTTCTGGTTAGGTACCGTCAAGGT GCCGCCCTATTTGAACGGCACTTGTTCTTCCCTAACAACAGAGCTTTACGATCCGAAAACCTTCATCACTCACGCG GCGTTGCTCCGTCAGACTTTCGTCCATTGCGGAAGATTCCCTACTGCTGCCTCCCGTAGGAGTCTGGGCCGTGTCT CAGTCCCAGTGTGGCCGATCACCCTCTCAGGTGGGCTACGCATCGTCGCCTTGGTGAGCCGTTACCTCACCAACTA GCTAATGCGCCGCGGGTCCATCTGTAAGTGGTAGCCGAAGCCACCTTTTATGTCTGAACCATGCGGTTCAAACAAC CATCCGGTATTAGCCCCGGTTTCCCGGAGTTATCCCAGTCTTACAGGCAGGTTACCCACGTGTTACTCACCCGTCC GCCGCTAACATCAGGGAGCAAGCTCCCATCTGTCCGCTCA。
The bacterial strain is bacillus amyloliquefaciens (Bacillus amyloliquefaciens), on September 26th, 2017 It is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (abbreviation CGMCC, address: Chaoyang District, Beijing City The institute 3 of North Star West Road 1, Institute of Microorganism, Academia Sinica), deposit number is CGMCC No.14741.
Embodiment 2
1, Bacillus amyloliquefaciens strain TJ1 fermentation process and its biological control agent preparation process
Seed liquor culture medium prescription: by mass percentage, beancake powder 3.00%, corn flour 4.00%, glucose 0.40%, fish meal 0.50%, potassium dihydrogen phosphate 0.03%, precipitated calcium carbonate 1.20%, ammonium sulfate 0.30%, sodium chloride 0.10%, remaining as water, pH is adjusted to 7.0-7.2.
Bulk fermentation culture medium prescription: with seed liquid culture medium formula.
Detailed process:
1. strain seed liquor culture: by Bacillus amyloliquefaciens strain TJ1 from test tube slant a small amount of bacterium of picking, move to In seed liquid culture medium, for 24 hours, this is seed liquor to 28 DEG C of shaking table shaken cultivations；
2. bulk fermentation culture: bulk fermentation culture medium material is sterilized (121~124 DEG C of temperature, 30 points in the fermenter Clock) aseptically, cultured seed liquor (inoculation volume ratio 10%) is poured into fermentor.28 DEG C of fermented and cultured 30h. Turn spore rate up to 80% or more, bacterium number is up to 80~9,000,000,000/g；
3. adding carrier (attapulgite powder) in the fermentation liquid of fermentation ends, it is uniformly mixed, the mass ratio with carrier: 1: 0.05,178 DEG C of spray-drying installation inlet air temperature, 170 DEG C of leaving air temp.Fermentation liquid through centrifugal spray to the 10% of original weight amount, Bacterium powder is obtained, wherein bacillus amyloliquefaciens TJ1 content >=75,000,000,000 viable bacterias/g；
4. taking each raw material by following weight parts: step 3. made bacillus amyloliquefaciens TJ1 bacterium powder 20%；Starch 8%；Portugal Grape sugar 10%；Lauryl sodium sulfate 8%；Pull open powder 2%；Bean cake powder 10%；Surplus is filled with attapulgite powder；
5. above-mentioned raw materials are uniformly mixed, the biological control agent of prevention and treatment celery root rot is obtained, wherein solution starch gemma bar Bacterium TJ1 content >=15,000,000,000 viable bacterias/g.
3 potting pharmacodynamic test of embodiment
The biological control agent for present embodiments providing the original powder of TJ1 containing Bacillus amyloliquefaciens strain is anti-to celery root rot Control the related experiment of effect.
1) reagent agent
The biocontrol agent (>=150CFU/g) of the bacterium powder of TJ1 containing Bacillus amyloliquefaciens strain, 70% fenaminosulf wettable Pulvis (Shanghai Ruilian Chemical Co., Ltd., commercially available)
2) for studying object and controlling object:
For studying object: celery (kind texts and pictures are drawn, and Shandong Rui You agricultural science and technology development corporation, Ltd. is commercially available)；
Controlling object: root rot.
3) experimental design and arrangement
Stem foot inocalation method is pierced using needle and mixes native insecticide-applying way, pot-culture method measurement is carried out to celery root rot control efficiency.
Medicament to be measured stirs evenly in mass ratio with substrate soil, 70% fenaminosulf wettable powder 1:800, the starch bud containing solution Biocontrol agent 1:800,1:1000,1:1200 of spore bacillus strain TJ1 bacterium powder.It is sub-packed in after mixing thoroughly in 8cm × 8cm pot for growing seedlings It is stand-by as drug containing matrix.
By upgrowth situation, consistent, seedling age is the celery seedling replanting of 35~40d into 8cm × 8cm drug containing substrate seedling alms bowl, Slow seedling 3d.Seedling-slowing stage is spent to seedling, celery is stabbed with the needle through 75% alcohol surface sterilization when erect plants, growth are normal and plants Strain basal part of stem 1~2 time, stabs but does not pierce, and it is 1 × 10 that concentration is applied in backward wound filling7The Fusarinm solani bacterium of CFU/ml Suspension 1ml.3 repetitions of every processing, 40 plants of celery seedling of every repetition.After artificial infection pathogen, by vegetable material 26~30 Moisturizing culture under conditions of DEG C with relative humidity 70%~85%.Wherein clear water processing only connects bacterium and is not administered, and blank control does not connect Bacterium is not administered.
A seedling incidence is observed every 8h after celery seedling inoculation, when clear water disease incidence is up to 65% or more, system Each processing plant incidence is counted, and calculates preventive effect.It calculates according to the following formula and manages disease incidence and control efficiency everywhere.
Disease incidence (%)=morbidity strain number/total plant amount × 100 of the processing
Control efficiency (%)=(clear water control group average attack rate-processing average attack rate)/clear water compares average onset Rate × 100
5) interpretation of result
(1) control efficiency of the reagent agent to celery root rot
Diseased plant number is investigated, for table 1 the results show that the diseased plant rate of clear water control is up to 70.13%, the diseased plant rate of each processing is equal It is compareed lower than clear water.The biological prevention and control agent and substrate soil weight ratio of the original powder of TJ1 containing Bacillus amyloliquefaciens strain are 1:800,1: 1000, treated that diseased plant rate is respectively 11.62%, 20.05%, 26.06% by 1:1200, at 70% fenaminosulf wettable powder The diseased plant rate of reason is 24.38%；The control efficiency of each processing of biological prevention and control agent of Bacillus amyloliquefaciens strain TJ1 original powder is distinguished It is 83.43%, 71.42%, 62.85%, wherein the control efficiency of 1:800 is best.Other processing have in the level of P < 0.05 Significant difference, 70% fenaminosulf wettable powder are 65.24% to the control efficiency of celery root rot.The starch gemma bar containing solution The harm of celery root rot can be effectively controlled in the biological prevention and control agent of bacteria strain TJ1 original powder.
The disease incidence and control efficiency of celery root rot after each processing of table 1
Note: with there is same letter to indicate that difference is not significant (P < 0.05) after Duncan Multiple range test after data line.
(2) celery security survey: it is observed that each chemicals treatment district is compared with check plot, celery growth is normal, no phytotoxicity It generates, illustrates the biological prevention and control agent of the original powder of TJ1 containing Bacillus amyloliquefaciens strain safe to celery for examination concentration.
Therefore, from the point of view of disease incidence and control efficiency, Bacillus amyloliquefaciens strain TJ1 has celery root rot preferable Control efficiency, can efficiently inhibit disease fungus conidia germination, mycelia growth, and be colonized in wound formed biomembrane, have Effect prevents disease fungus from infecting celery.Control efficiency is up to 80% or more after application, hence it is evident that is better than comparison medicament, difference is aobvious It writes.
SEQUENCE LISTING
<110>Qingdao Calendula Bell bioengineering Co., Ltd
<120>one bacillus amyloliquefaciens and its application in prevention and treatment celery root rot
<130> 0
<160> 1
<170> PatentIn version 3.3
<210> 1
<211> 1400
<212> DNA
<213>the 16s rDNA gene order of bacillus amyloliquefaciens (Bacillus amyloliquefaciens) Tj1
<400> 1
ctagaggtta ccttaccgac ttcgggtgtt acaaactctc gtggtgtgac gggcggtgtg 60
tacaaggccc gggaacgtat tcaccgcggc atgctgatcc gcgattacta gcgattccag 120
cttcacgcag tcgagttgca gactgcgatc cgaactgaga acagatttgt gggattggct 180
taacctcgcg gtttcgctgc cctttgttct gtccattgta gcacgtgtgt agcccaaggt 240
ataagggggc atgatgattt gacgtcatcc ccaccttcct ccggtttgtc accggcagtc 300
accttagagt gcccaactga atgctggcaa ctaagatcaa gggttgcgct cgttgcggga 360
cttaacccaa catctcacga cacgagctga cgacaaccat gcaccacctg tcactctgcc 420
cccgaagggg acgtcctatc tctaggattg tcagaggatg tcaagacctg gtaaggttct 480
tcgcgttgct tcgaattaaa ccacatgctc caccgcttgt gcgggccccc gtcaattcct 540
ttgagtttca gtcttgcgac cgtactcccc aggcggagtg cttaatgcgt tagctgcagc 600
actaaggggc ggaaaccccc taacacttag cactcatcgt ttacggcgtg gactaccagg 660
gtatctaatc ctgttcgctc cccacgcttt cgctcctcag cgtcagttac agaccagaga 720
gtcgctttcg ccactggtgt tcctccacat ctctacgcat ttcaccgcta cacgtggaat 780
tccactctcc tcttctgcac tcaagttccc cagtttccaa tgaccctccc cggttgagcc 840
gggggctttc acatcagact taagaaaccg cctgcgagcc ctttacgccc aataattccg 900
gacaacgctt gccacctacg tattaccgcg gctgctggca cgtagttagc cgtggctttc 960
tggttaggta ccgtcaaggt gccgccctat ttgaacggca cttgttcttc cctaacaaca 1020
gagctttacg atccgaaaac cttcatcact cacgcggcgt tgctccgtca gactttcgtc 1080
cattgcggaa gattccctac tgctgcctcc cgtaggagtc tgggccgtgt ctcagtccca 1140
gtgtggccga tcaccctctc aggtgggcta cgcatcgtcg ccttggtgag ccgttacctc 1200
accaactagc taatgcgccg cgggtccatc tgtaagtggt agccgaagcc accttttatg 1260
tctgaaccat gcggttcaaa caaccatccg gtattagccc cggtttcccg gagttatccc 1320
agtcttacag gcaggttacc cacgtgttac tcacccgtcc gccgctaaca tcagggagca 1380
agctcccatc tgtccgctca 1400

Claims (6)

  1. Bacillus amyloliquefaciens 1. (Bacillus amyloliquefaciens) TJ1, the deposit number of the bacterial strain is CGMCC No.14741。
  2. 2. purposes of the Bacillus amyloliquefaciens strain TJ1 described in claim 1 in prevention and treatment celery root rot.
  3. 3. a kind of biological control agent, including Bacillus amyloliquefaciens strain TJ1 described in claim 1.
  4. 4. the preparation method of biological control agent as claimed in claim 3, characterized in that the following steps are included:
    (1) seed liquor of Bacillus amyloliquefaciens strain TJ1 is prepared；
    (2) seed liquor prepared by step (1) is inoculated into bulk fermentation culture medium, constant temperature incubation 28h-32h at 26-30 DEG C；
    (3) carrier will be added in the fermentation liquid of step (2) culture, spray drying obtains bacillus amyloliquefaciens TJ1 bacterium powder；
    (4) bacillus amyloliquefaciens TJ1 bacterium powder is added auxiliary agent and biological control agent is made.
  5. 5. the preparation method of biological control agent as claimed in claim 4, characterized in that
    The seed liquid culture medium that the preparation seed liquor uses are as follows: by mass percentage, beancake powder 3.00%, corn flour 4.00%, glucose 0.40%, fish meal 0.50%, potassium dihydrogen phosphate 0.03%, precipitated calcium carbonate 1.20%, ammonium sulfate 0.30%, sodium chloride 0.10%, remaining as water, pH is adjusted to 7.0-7.2；
    Bulk fermentation culture medium: with seed liquid culture medium.
  6. 6. purposes of the biological control agent as claimed in claim 3 in prevention and treatment celery root rot.
CN201811052395.4A 2018-09-10 2018-09-10 One bacillus amyloliquefaciens and its application in prevention and treatment celery root rot Withdrawn CN109207393A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111139199A (en) * 2020-01-13 2020-05-12 中化农业（临沂）研发中心有限公司 Bacillus amyloliquefaciens CGMCC No.17843 and application thereof
CN113303340A (en) * 2021-05-21 2021-08-27 宁夏大学 Composition of biocontrol bacteria and fish protein and application of composition in cucumber cultivation

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111139199A (en) * 2020-01-13 2020-05-12 中化农业（临沂）研发中心有限公司 Bacillus amyloliquefaciens CGMCC No.17843 and application thereof
CN111139199B (en) * 2020-01-13 2021-08-31 中化农业（临沂）研发中心有限公司 Bacillus amyloliquefaciens CGMCC No.17843 and application thereof
CN113303340A (en) * 2021-05-21 2021-08-27 宁夏大学 Composition of biocontrol bacteria and fish protein and application of composition in cucumber cultivation

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