CN108048354A - One bacillus subtilis and its application - Google Patents
One bacillus subtilis and its application Download PDFInfo
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Abstract
The present invention provides a bacillus subtilis (Bacillus subtilis) LX13, and deposit number is CGMCC No.8805.Above-mentioned bacillus subtilis LX13 can be used for preventing in peanut crown rot, southern blight and root rot.Compared with prior art, bacillus subtilis LX13 provided by the invention has apparent antagonism to peanut pathogen, can be used as potentiality biocontrol bacterial strain；Biocontrol effect is good, has apparent control effect to crown rot, southern blight and root rot；Growth and the yield of peanut can be obviously promoted；It is easy to use, rapid-action, long action time, easily prepared and preservation, convenient for applying and promoting.
Description
Technical field
The invention belongs to microbial technology fields, and in particular to a kind of bacillus subtilis and its application.
Background technology
Peanut crown rot is a kind of peanut time of infertility generable soil-borne fungus type disease, and cause of disease is
Aspergillus nigerV.Tiegh belong to Deuteromycotina, hyphomycetales Eurotium, black-koji mould fungi.The disease is mainly sent out
It gives birth in seedling stage or grows early period.In peanut seedling stage, germ first infects the cotyledon of remaining, and then the basal part of stem of infecting peanut.Sick portion
The concave scab of nascent yellowish-brown, scab edge brown, later disease portion expand rapidly, and cortex lobe organizes dry rot, finally only surplus
Broken fibr tissue down.In the case of humidity, sick portion grows the mould layer of black quickly, i.e., the mitogenetic spore of germ is in stalk and mitogenetic
Spore.Kernel is infected, can be allowed to rot and cannot germinate, one layer of black mould is grown in aggrieved portion faces.After benevolence germination, disease
Bacterium can infect cotyledon, and cotyledon is not unearthed and rots with regard to blackening, and it is in water soaking mode that plumular axis, which is infected, light brown, there is the mould layer of black.Peanut
Growth period is susceptible, often results in basal part of stem and rots, and diseased plant is wilted withered.
Peanut sclerotium rolfsii is the important disease on peanut, and cause of disease is Sclerotium rolfsii Sacc., belongs to Fungi Imperfecti
Subphylum, no spore Zoopagales, Sclerotium rolfsii category fungi.The disease mostly occurs in Adult plant, and hot and humid 7~September part is that morbidity is contained
Phase, main harm basal part of stem, carpopodium, fruit pod and root.Aggrieved diseased tissues initial stage is in the soft corruption of crineous, grows white spun silk soon
The mycelia of shape is covered by sick position.Environmental condition suitable for when, mycelia outwards spreads rapidly, the cane of peanut middle and lower part near the ground
And the soil surface around diseased plant, it can all grow one layer of bombycine subiculum of white；Later stage is formed in sick portion's subiculum
Many sclerotium；With rotting for aggrieved diseased tissues, plant cortex comes off, only remaining fibr tissue, very frangibility.After pod is susceptible
Sick portion becomes light brown to crineous, becomes shrinkage after kernel is susceptible and rots, sick portion covers taupe subiculum, and the later stage can also form bacterium
Core；Germ inside kind of shell and benevolence surface growth when can also generate oxalic acid so that on kind of skin formed striped, sheet or
Circular black-and-blue lathe work.
Peanut root rot can occur in peanut each breeding time, and cause of disease is Deuteromycotina Fusarium fungi.Before emergence
Rotten kind, rotten bud can be caused by catching an illness；It is aggrieved in seedling stage to cause seedling withered；Cause root-rot, brown foot rot and pod rotten Adult plant is aggrieved,
The short and small, undergrowth of diseased plant overground part performance, blade from bottom to top dry up after gradual change Huang, main root browning, shrinkage, dry rot, lateral root
It comes off or lateral root is few and short, main root only stays the root tissue of remaining, be commonly called as its " mouse tail " as mouse tail when pulling up.
Currently to the prevention of peanut pest and disease damage mainly based on plant resistance to environment stress kind and chemical prevention, but Pest-resistant
Problem getting worse, and chemical pesticide residual causes serious harm, and such as residual hazard, environmental pollution, human body and ecology are caused
Very big threat, is unfavorable for the sustainable development of peanut cultivation industry.Therefore, a kind of economic, safe and effective control measure is found
It is extremely urgent.Bio-control method is to be killed or reduced the quantity of causal organism using beneficial microbe to control plant disease
Occur, a kind of measure of development.These beneficial organisms are also known as antagonistic microbe or biocontrol microorganisms.It is related to peanut biocontrol microorganisms at present
Research, still, for above-mentioned three cultivate peanut pathogen biocontrol agent in application process there are preventive effect it is unstable, work it is slow,
The problem of long action time, single preventive and therapeutic effect so that the application of microbial bacterial agent receives obstruction.
The content of the invention
In view of the above-mentioned problems, the present invention is intended to provide a kind of bacillus subtilis (Bacillus subtilis), acts on
Peanut rhizosphere has the effect of extraordinary prevention peanut crown rot, southern blight and root rot.
One bacillus subtilis (Bacillus subtilis) LX13, deposit number are CGMCC No.8805.
Applications of the above-mentioned bacillus subtilis LX13 in prevention peanut crown rot, southern blight and root rot.
Further, the application is specially pouring root after the bacterium solution of bacillus subtilis LX13 is diluted.
Bacillus subtilis LX13 is prepared into the microbial inoculum of prevention peanut crown rot, southern blight and root rot.
Applications of the above-mentioned bacillus subtilis LX13 in terms of peanut growth and output increased is promoted.
Further, pouring root after the bacterium solution of careless bacillus LX13 is diluted.
Compared with prior art, bacillus subtilis LX13 provided by the invention has peanut pathogen apparent antagonism to make
With potentiality biocontrol bacterial strain can be used as；Biocontrol effect is good, has apparent control effect to crown rot, southern blight and root rot；Energy
Enough it is obviously promoted growth and the yield of peanut；It is easy to use, rapid-action, long action time, easily prepared and preservation, convenient for answering
With and promote.
Description of the drawings
Fig. 1 is the colonial morphology figure of bacterial strain LX13；
Fig. 2 is the PCR product amplification figure of the 16s rDNA of bacterial strain LX13；
Fig. 3 be bacterial strain LX13 with its in GenBank databases it is related belong to plant build based on 16S rDNA sequences
Phylogenetic tree.
Biological material specimens preservation information：
Bacillus subtilis (Bacillus subtilis) LX13, is preserved in Chinese microorganism strain preservation conservator
Meeting common micro-organisms center (CGMCC), preservation address：China, the institute 3 of Chaoyang District Beijing North Star West Road 1, the Chinese Academy of Sciences is micro-
Biological study institute, preservation date：On 01 22nd, 2014, deposit number CGMCC No.8805.
Specific embodiment
The present invention is described in further details with reference to specific embodiment and attached drawing.
The material of following embodiment is as follows：
Isolation medium：NA culture mediums：Beef extract (Beef extract) 3g, peptone (Peptone) 5g, glucose
(Dextrose) 10g, agar powder (Agar) 15g, distilled water 1000mL adjust pH to 7.0,121 DEG C of high pressure steam sterilization 20min.
Seed culture medium：LB fluid nutrient mediums, ingredient：Tryptone (Tryptone) 10g, yeast extract (Yeast
Extract) 5g, sodium chloride (NaCl) 10g, distilled water 1000mL adjust pH to 7.0,121 DEG C of high pressure steam sterilization 20min.
Fungi culture medium：PDA culture medium, ingredient：Potato 200g, glucose 20g, agar powder 15g, distilled water
1000mL, 121 DEG C of high pressure steam sterilization 20min.
Test originated from fungus：Shandong Peanut Inst. preserves peanut crown rot, sclerotium rolfsii and peanut pine root fungus.
Separation, culture and the identification of 1 bacterial strain of embodiment
1st, the screening and separation of bacterial strain
A collection of single bacterium on picking tablet falls within setting-out on LB solid medium tablets and is inverted in 28 DEG C of constant incubator trainings
It supports 1-2 days.
Soil comes from Shandong Peanut Inst.'s experimental field peanut rhizosphere soil.Weighing 5g rhizosphere soils, to be put into 45ml sterile
In the triangular flask of water, a moment is shaken, upper liquid 0.5ml is drawn and is added to containing in 4.5ml sterile water test tubes, according to 102、103、
104、105、106Multiple carry out gradient dilution.
The 100 μ l of soil supension of different extension rates are drawn respectively with micropipettor in the spreader that on NA tablets, sterilizes
Coating hooks, and is inverted in after tablet is sealed in 28 DEG C of constant incubators and cultivates 48h.
A collection of single bacterium on picking tablet falls within setting-out on LB solid medium tablets and is inverted in 28 DEG C of constant incubator trainings
Support 1-2d.
2nd, the Physiology and biochemistry of the morphologic observation of bacterium colony and bacterial strain is identified
Biocontrol bacteria bacterial strain LX13 is grown on LB tablets, milky, and dry tack free has gauffer, matt, rough, bacterium
Fall edge to be irregularly serrated, as shown in Figure 1；Bacterial strain LX13 is in clear LB fluid nutrient mediums, and shaken cultivation, surface is not
Film layer is formed, bacteria suspension is muddy.
The ginsengs such as Gram's staining, V-P measure, catalase test, nitrate reduction, citrate utilization, methyl red test
According to《Common bacteria system identification handbook》Carry out Physiology and biochemistry identification.
The Physiology and biochemistry qualification result of LX13 bacterial strains is shown in Table 1, and the results show LX13 thalline show as Gram-positive, aerobic
Bacterium；Methyl red, catalase and V-P reactions are positive；Orthonitric acid can be gone back using citrate as the source of nutrition of carbon
Salt, caseinhydrolysate, starch and gelatin；And sugar alcohol can be fermented and generate acid, but edwardsiella hoshinae.According to more than morphological feature and
Physio-biochemical characteristics, and combine primary Jie Shi Bacteria Identifications handbook and common bacteria system identification handbook, biocontrol bacteria bacterial strain LX13
It is sufficiently close to bacillus subtilis (Bacillus subtilis).
The physiological and biochemical property of 1 active bacterial strain LX13 of table
Note："+" represents reaction result for the positive；"-" represents reaction result for feminine gender.
3rd, the 16S rRNA molecules identification of bacterium
Bacterial strain DNA is extracted
The bacterium bacterial strain that there is antagonistic activity to fungi is chosen, 10 identical bacterium colonies of picking do repetition, carry out genome
The extraction of DNA, extracting method are carried out referring especially to TIANGEN TIANamp BACTERia DNA Kit kit specifications,
And change slightly, step is as follows：
(1) inoculum 1mL, 10000rpm centrifugation 1min is taken, exhaust supernatant as far as possible；
(2) 200 μ L buffer solution GA are added in into bacterial sediment, shakes to thalline and thoroughly suspends；
(3) 4 μ LRNAase (100mg/mL) solution are added in, 15s is shaken, is placed at room temperature for 5min；
(4) 20 μ L Proteinase K Solutions, mixing are added in into pipe；
(5) 220 μ L buffer solution GB are added in, shake 15s, 70 DEG C of placement 10min, brief centrifugation is to remove the water of cap wall
Pearl；
(6) 220 μ L absolute ethyl alcohols are added in, fully shaking mixing 15s is likely to occur flocculent deposit at this time, brief centrifugation with
Remove the droplet of cap wall；
(7) solution obtained by previous step and flocculent deposit all add in an adsorption column GB3 to (adsorption column is put into collecting pipe
In), 12000rpm centrifugation 30s, outwelling waste liquid will be put into adsorption column CB3 in collecting pipe；
(8) 500 μ L buffer solutions GD (whether preoperation inspection is added into absolute ethyl alcohol) are put into adsorption column CB3,
12000rpm centrifuges 30s, and outwelling waste liquid will be put into adsorption column CB3 in collecting pipe；
(9) 700 μ L rinsing liquids PW (whether preoperation inspection is added into absolute ethyl alcohol) are put into adsorption column CB3,
12000rpm centrifuges 30s, and outwelling waste liquid will be put into adsorption column CB3 in collecting pipe；
(10) 500 μ L rinsing liquids PW, 12000rpm centrifugation 30s are put into adsorption column CB3, outwell waste liquid by adsorption column
It is put into CB3 in collecting pipe；
(11) adsorption column CB3 is put back in collecting pipe, 12000rpm centrifugation 2min outwell waste liquid, adsorption column CB3 is placed in
It is placed at room temperature for several minutes, thoroughly to dry rinsing liquid remaining in sorbing material；
(12) adsorption column CB3 is transferred in clean centrifuge tube, 50-200 μ L is vacantly added dropwise to the middle part of adsorbed film
Elution buffer TE, is placed at room temperature for 2-5min, and solution is collected into centrifuge tube, -20 DEG C of preservations by 12000rpm centrifugation 2min.
PCR and sequencing
(1) DNA of extraction is with reference to the 16s rDNA Bacterial Identification PCR Kit of TaKaRa companies
Kit specification carries out PCR amplification, and wherein forward primer is：5’-AGAGTTTGATCATGGCTCAG-3’(SEQ ID
No.1), reverse primer is：3’-CGCTTACCTTGTTACGACTT-5’(SEQ ID No.2).
PCR amplification condition is as follows：94 DEG C of pre-degeneration 5min；94 DEG C of denaturation 1min；53 DEG C of annealing 1min；72 DEG C of extension 90s；
Extend 5min after 72 DEG C, totally 30 Xun Huans.
PCR product is separated using 1% agarose gel electrophoresis, and EB dyeing is placed on 3UVTMTransilluminator
It is observed under (UVP, USA).Find that LX13 bacterial strain 16s rDNA clip sizes are about 1500bp by electrophoresis detection, segment is big
Small to be consistent with kit desired design, amplification is shown in Fig. 2.
(2) according to the primer being pre-designed and expected amplified fragments size, binding marker (Marker) target fragment is in purple
It is cut down under outer lamp, the recycling of DNA is carried out with QIAquick Gel Extraction Kit Silica Bead DNA GelExtraction Kit.16s
RDNA sequencings carry out sequencing by precious bioengineering (Dalian) Co., Ltd.It measures sequence results and shows LX13 bacterial strains
16srDNA segments share 1456nt base-pairs composition, and sequence table information is shown in SEQ ID No.3.By blast program by measure
Sequence and GenBank (NCBI, website http://blast.ncbi.nlm.nih.gov/Blast.cgi) in sequence compare,
Then obtain from GenBank and be determined with test strain Sequences similar kind, the 16s rDNA sequences belonged to.Comparison result is shown in figure
3, the bacillus subtilis of bacillus in the 16s rDNA sequence GenBank gene pools of the results show active bacterial strain
(Bacillussubtilis) 16s rDNA sequence very high homologies, homology is up to 99%.Structure development tree and homology analysis
Bacillus subtilis (Bacillus subtilis) (Accession of the results show (Fig. 3), bacterial strain LX13 and bacillus
No:EF47226.1) it is separately formed a branch, closest in evolution, affiliation is nearest between reflecting them, fortune
It is 99% to analyze to obtain the two homology with DANMAN softwares.With reference to traditional physio-biochemical characteristics identification and 16S rDNA sequences
Arrange analyzing as a result, judging that bacterial strain LX13 is bacillus subtilis.
2 bacterial strain of embodiment inhibits the measure of fungi ability
Peanut crown rot, sclerotium rolfsii and peanut pine root fungus are taken into about 0.5cm* with tweezers in aseptic operating platform
The square tiles of 0.5cm are seeded in PDA culture medium, and culture 2-3 days is inverted in 28 DEG C of greenhouses.
The picking single bacterium colony LX13 best to fungi restraining effect carries out fungi face-off experiment.Treat that fungi is grown to accounting for cultivating
In the inoculated bacteria at fungi 1-2cm during 1/3 size of ware, continue 28 degrees Celsius of placement hot-house culture 2-3 days, observation fungi life
Long situation.
Bacteriostasis rate=[(control fungi growth radius-processing fungi growth radius)/control fungi growth radius] × 100%
Experimental result：LX13 carries out the dual test of peanut crown rot bacterium 3 times altogether, is shown in Table 2, and inhibitory action is apparent, most
Low inhibiting rate is minimum 75.72%, up to 77.15%.LX13 carries out the dual test of peanut sclerotium rolfsii 3 times altogether, sees
Table 3, inhibitory action is apparent, and minimum inhibiting rate is minimum 75.44%, and up to 76.09%.LX13 is to the 3 of peanut root rot
Secondary dual test, is shown in Table 4, minimum to the inhibition of peanut root rot 66.82%, up to 69.60%.It is indicated above that
LX13 has apparent antagonism to peanut pathogen, can be used as potentiality biocontrol bacterial strain.
Table 2LX13 and the face-off experimental result (three batches of experiments, every time three repetitions) of peanut crown rot bacterium
The face-off experimental result of table 3LX13 and Sclerotium rolfssi (three batches of experiments, every time three repetitions)
The face-off experimental result (three batches of experiments, every time three repetitions) of table 4LX13 and peanut pine root fungus
3 bacterial strain indoor pot cure plant disease of peanut of embodiment and growth-promoting functions experiment
LB fluid nutrient mediums are accessed after bacterial strain is activated, are placed in 180r/min shake cultures 3d in 28 degree of shaking tables.
The expansion of the pathogen of the peanut crown rot of preservation, southern blight and root rot is numerous：Oat grain is fitted into triangular flask,
Moisture, 121 DEG C of high pressure sterilization 20min are outwelled after distilled water immersion 6h；By peanut crown rot bacterium, sclerotium rolfsii, pine root fungus point
It is not inoculated into the oat grain of sterilizing, is placed in 28 DEG C of culture 7d, is shaken every day inoculation bottle 2 times, all oat grains is made to have cause of disease
Bacterium obtains the oat grain that carries disease germs.
Pathogen is inoculated with：By peanut (spending No. 8 in kind Shandong) plantation in the basin of phjytotron, soil：Vermiculite：Nutrition Soil
Mass ratio=2：1：1.Cultivation temperature daytime is 28 DEG C, and at 25 DEG C of night, 12h illumination/12h is dark, peanut simple grain plantation, per basin
3.10 utilizations of wheat of carrying disease germs of each pathogen spread table local method and are inoculated in peanut periphery, and subsequent lid last layer thin soil simultaneously waters.
LX13 is inoculated with：Respectively sowing time, after planting 10d utilize bacillus subtilis fermentation liquor pouring root, each handle 10
Basin per 3 plants of basin, is repeated 3 times.Pour 100 times of zymotic fluid 100m L of dilution every time per basin.50% carbendazim, 800 times of liquid and clear are set
Two controls of water.The morbidity feelings of crown rot, southern blight and root rot are investigated in observation peanut incidence daily, plantation after 75 days
Condition.Peanut watering 1 time daily.During harvesting peanut, influences of the investigation LX13 to Development of Peanut.
Crown rot grade scale：0 grade：Plant is asymptomatic；1 grade：Only scab is generated in basal part of stem；2 grades：Basal part of stem generation is hung
Contracting symptom, less than 1/3rd performance systemic symptoms (withered, dead, wilting etc.) of whole strain；3 grades：2/3rds of whole strain with
Lower performance systemic symptom；4 grades：More than 2/3rds representation system symptoms of whole strain, dead plant is by 4 grades of calculating.
Southern blight grade scale：0 grade：Plant is asymptomatic；1 grade：Scab is only generated in basal part of stem；2 grades：Basal part of stem generation is hung
Contracting symptom, the affected part of representation system symptom (withered, dead, wilting etc.) account for less than 1/3rd of whole strain；3 grades：Performance
The affected part of systemic symptom accounts for less than 2/3rds of whole strain；4 grades：The affected part of representation system symptom accounts for three points of whole strain
More than two.
Root rot grade scale：0 grade:Equal disease-free spot on stem foot and main fibrous root；1 grade:There is a small amount of scab on stem foot and main root；
3 grades:Scab is more on stem foot and main root, and lesion area accounts for the 1/4~1/2 of stem foot and the root gross area；5 grades:On stem foot and main root
Scab is more and big, and lesion area accounts for the 1/2~3/4 of stem foot and the root gross area；7 grades:On stem foot and main root scab in flakes, formed around
Stem phenomenon, but root system is not dead；9 grades:Root system necrosis, plant above ground portion wilts or death.
Diseased plant rate=morbidity strain number/total strain number × 100%
Disease index=∑ (morbidity grade typical value × diseased plant number at different levels) × 100/ (investigation total strain number × superlative degree morbidity generation
Tabular value)
Control effect=[(control disease index-processing disease index)/control disease index] × 100%
Potted plant experiment result：With the fermentation liquor treatment peanut bacterial strain of biocontrol microorganisms LX13, LX13 is preced with indoor pot peanut rotten
The control effect of disease is 67.5%, is shown in Table 5；LX13 is 57.6% to the control effect of indoor pot peanut sclerotium rolfsii, is shown in Table 6；
LX13 is 61.2% to the control effect of indoor pot peanut root rot, is shown in Table 7, is above the processing of comparison medicament carbendazim.This
Illustrate that biocontrol microorganisms LX13 has apparent control effect to peanut crown rot, southern blight and root rot.
5 bacterial strain LX13 of table is to the control effect of peanut crown rot
6 bacterial strain LX13 of table is to the control effect of peanut sclerotium rolfsii
7 bacterial strain LX13 of table is to the control effect of peanut root rot
Peanut seeding is harvested after 4 months, pours out potting peanut, measurement peanut stem height, side shoot length, ground diameter, branch
Number；Peanut pod number is recorded, naturally dry and is weighed after washing pod surface soil particle with flowing water.Peanut is cut off with scissors
Plant above ground, under ground portion are put into after drying to constant weight in drying box at 80 DEG C and weigh its dry weight.Experimental result is shown in Table 8.
Table 8LX13 is inoculated with the influence to peanut growth
Measurement peanut stem is high at 120 days after peanut seeding, side shoot is long, ground diameter is thick, branch amount, finds to be inoculated with LX13 peanuts
Average stem is high, side shoot is long, ground diameter is thick, branch amount is above not being inoculated with peanut and carbendazim medicament processing.Connect bacterium peanut single plant
Pod number, pod weight, overground part dry weight, underground part dry weight are also all higher than not being inoculated with peanut and carbendazim medicament processing, this says
Bright, inoculation LX13 significantly facilitated growth and the yield of peanut.
4 bacterial strain LX13 control in field peanuts crown rot of embodiment, southern blight and root rot experiment
Experiment is carried out in Shandong Peanut Inst. Laixi experimental plot, for peanut continuous cropping field, peanut crown rot, southern blight
It is serious with the generations such as root rot.Using RANDOMIZED BLOCK DESIGN, peanut is per cell 70m2, 4 repetitions.Clear water is blank control,
50% 800 times of carbendazim liquid is positive control.Respectively sowing time, after planting 10d using LX13 ferment liquid irrigating root, every plant every time
Pour dilution 100 times of zymotic fluids about 100m L.Other field management investigate incidence with normal production after 75 days.
Field experiment the result shows that, the fermentation liquor treatment peanut bacterial strain of Field information biocontrol microorganisms LX13, LX13 to peanut be preced with
The control effect of maize ear rot is 62.5%, is shown in Table 9；LX13 is 52.0% to the control effect of peanut sclerotium rolfsii, is shown in Table 10；LX13 pairs
The control effect of peanut root rot is 56.1%, is shown in Table 11.This illustrates biocontrol microorganisms LX13 to peanut crown rot, southern blight and root-rot
Disease has apparent control effect, and to crown rot, root rot control effect due to carbendazim processing.
9 Field information bacterial strain LX13 of table is to the control effect of peanut crown rot
10 Field information bacterial strain LX13 of table is to the control effect of peanut sclerotium rolfsii
11 Field information bacterial strain LX13 of table is to the control effect of peanut root rot
Before harvesting peanut, each processing takes 30m2It dries and weighs, calculate yield, experimental result is shown in Table 12.LX13 processing
Average product is converted into per mu yield as 239.72kg, is compareed more than comparison medicament carbendazim (217.00kg) and clear water
(129.22kg), than comparison medicament carbendazim volume increase more than 25.89%, than clear water control volume increase 85.52%.
Influences of the 12 Field information bacterial strain LX13 of table to peanut yield
To sum up, show it to hat with the fermentation liquor treatment peanut bacterial strain of biocontrol microorganisms LX13, indoor pot and field experiment
Maize ear rot, southern blight and root rot have apparent control effect, and significantly improve the yield of peanut.
Sequence table
<110>Shandong Peanut Inst.
<120>One bacillus subtilis and its application
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<170> SIPOSequenceListing 1.0
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<213>Artificial sequence (Artificial Sequence)
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cgcttacctt gttacgactt 20
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<213>Bacillus subtilis (Bacillus subtilis)
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taactccggg aaaccggggc taataccgga tggttgtttg aaccgcatgg ttcaaacata 180
aaaggtggct tcggctacca cttacagatg gacccgcggc gcattagcta gttggtgagg 240
taacggctca ccaaggcgac gatgcgtagc cgacctgaga gggtgatcgg ccacactggg 300
actgagacac ggcccagact cctacgggag gcagcagtag ggaatcttcc gcaatggacg 360
aaagtctgac ggagcaacgc cgcgtgagtg atgaaggttt tcggatcgta aagctctgtt 420
gttagggaag aacaagtacc gttcgaatag ggcggtacct tgacggtacc taaccagaaa 480
gccacggcta actacgtgcc agcagccgcg gtaatacgta ggtggcaagc gttgtccgga 540
attattgggc gtaaagggct cgcaggcggt ttcttaagtc tgatgtgaaa gcccccggct 600
caaccgggga gggtcattgg aaactgggga acttgagtgc agaagaggag agtggaattc 660
cacgtgtagc ggtgaaatgc gtagagatgt ggaggaacac cagtggcgaa ggcgactctc 720
tggtctgtaa ctgacgctga ggagcgaaag cgtggggagc gaacaggatt agataccctg 780
gtagtccacg ccgtaaacga tgagtgctaa gtgttagggg gtttccgccc cttagtgctg 840
cagctaacgc attaagcact ccgcctgggg agtacggtcg caagactgaa actcaaagga 900
attgacgggg gcccgcacaa gcggtggagc atgtggttta attcgaagca acgcgaagaa 960
ccttaccagg tcttgacatc ctctgacaat cctaggagat aggacgtccc ctttcggggg 1020
cagagtgaca ggtggtgcat ggttgtcgtc agctcgtgtc gtgagatgtt gggttaagtc 1080
ccgcaacgag cgcaaccctt gatcttagtt gccagcattc agttgggcac tctaaggtga 1140
ctgccggtga caaaccggag gaaggtgggg atgacgtcaa atcatcatgc cccttatgac 1200
ctgggctaca cacgtgctac aatggacaga acaaagggca gcgaaaccgc gaggttaagc 1260
caatcccaca aatctgttct cagttcggat cgcagtctgc aactcgactg cgtgaagctg 1320
gaatcgctag taatcgcgga tcagcatgcc gcggtgaata cgttcccggg ccttgtacac 1380
accgcccgtc acaccacgag agtttgtaac acccgaagtc ggtgaggtaa ccttttagga 1440
gccagccgcc gaag 1454
Claims (6)
1. a bacillus subtilis (Bacillus subtilis) LX13, deposit number is CGMCC No.8805.
2. applications of the bacillus subtilis LX13 described in claim 1 in prevention peanut crown rot, southern blight and root rot.
3. application according to claim 2, which is characterized in that the application is specially by the bacterium of bacillus subtilis LX13
Pouring root after liquid dilution.
4. application according to claim 2, which is characterized in that it is rotten that bacillus subtilis LX13 is prepared into prevention peanut hat
The microbial inoculum of disease, southern blight and root rot.
5. applications of the bacillus subtilis LX13 described in claim 1 in terms of peanut growth and output increased is promoted.
6. application according to claim 5, which is characterized in that pouring root after the bacterium solution of careless bacillus LX13 is diluted.
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| CN109122722A (en) * | 2018-07-24 | 2019-01-04 | 江苏师范大学 | Garlic endophyte bacillus subtilis is promoting the application in Garlic Growth |
| CN110810440A (en) * | 2019-11-13 | 2020-02-21 | 河南农贝得农业科技有限公司 | Microbial agent for preventing and treating crop southern blight and preparation method thereof |
| CN111602676A (en) * | 2020-07-03 | 2020-09-01 | 天津坤禾生物科技集团股份有限公司 | Application of bacillus subtilis MES810 in prevention and treatment of grape southern blight root rot |
| CN112322559A (en) * | 2020-12-09 | 2021-02-05 | 河南省科学院生物研究所有限责任公司 | Peanut endophytic bacterium bacillus subtilis F-1 and application thereof |
| CN112458008A (en) * | 2020-11-17 | 2021-03-09 | 山东省花生研究所 | Biocontrol bacillus subtilis ZHX-1 and application thereof |
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| CN111602676A (en) * | 2020-07-03 | 2020-09-01 | 天津坤禾生物科技集团股份有限公司 | Application of bacillus subtilis MES810 in prevention and treatment of grape southern blight root rot |
| CN111602676B (en) * | 2020-07-03 | 2021-09-07 | 天津坤禾生物科技集团股份有限公司 | Application of bacillus subtilis MES810 in prevention and treatment of grape southern blight root rot |
| CN112458008A (en) * | 2020-11-17 | 2021-03-09 | 山东省花生研究所 | Biocontrol bacillus subtilis ZHX-1 and application thereof |
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