CN108949641A - One bacillus pumilus bacterial strain and its application - Google Patents
One bacillus pumilus bacterial strain and its application Download PDFInfo
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Abstract
The invention discloses a bacillus pumilus bacterial strains, bacillus pumilus (Bacillus pumilus) bacterial strain is preserved in Guangdong Province's Culture Collection, it is named as bacillus pumilus CFC-5 (Bacillus pumilus CFC-5), it is GDMCC No:60397 in the deposit number of Guangdong Province's Culture Collection, and the preservation time is on June 25th, 2018.Strain of i (bacillus) pumilus provided by the invention is bacillus pumilus CFC-5, which has the ability of protein in degradation tung oil tree grouts, has wide spectrum inhibiting effect to crops soil-borne disease, and have growth-promoting functions, provides bacterium source for degradation tung oil tree grouts.The invention also discloses application of the bacterial strain in degradation tung oil tree grouts in protein and inhibition corps diseases.
Description
Technical field
The invention belongs to microbial engineering field more particularly to a bacillus pumilus bacterial strain and its applications.
Background technique
Tung oil tree is the woody oil tree species in China native country, has fast growth, yield height, wide adaptability, seed oil content high
And the advantages that full of nutrition.Paulownia cake after the oil expression of paulownia seed, contains nitrogen 4%~6%, P2O51.8%~2.7%, K2O 1.2%~
1.3%, it to be a kind of excellent protein resource that there are also a large amount of other microelements.But since tung oil tree grouts contain saponin and Buddhist
The toxin such as wave alcohol and its related chemical species, thus tung oil tree grouts are mostly used as cake fertilizer returning to the field for a long time, cannot function as moving
Object food source.
In agricultural production, high nutrient and simplicity fertilizer application it is excessive, bring it is various agricultural and environmental problems: soil
Nutrient loss, environmental pollution, crop disease-resistant insect pest reduced capability, wasting of resources etc..Paulownia dregs of rice fermentation after processing is prepared into
Bio-bacterial manure not only facilitates the diversity for solving bio-fertilizer, and can evade to a certain extent for chemical fertilizer
It relies on, can also reduce the use of pesticide.But tung oil tree grouts directly applied as fertilizer or heap fermentation after apply, it is not only fertile
Effect is slow, and the nutriment in the paulownia dregs of rice cannot make full use of and result in waste of resources.
Bacillus pumilus (Bacillus pumilus) is a kind of Gram positive aerobic bacillus, be can produce
Gemma.Bacillus pumilus can very strong extracellular proteinase, cellulase, lipase, zytase etc., be conducive to degraded oil
Macromolecular substances in paulownia grouts can also generate the antagonistic substances such as antibiotic and antibacterial protein, and scope of restraining fungi is wide.Such as: Yin
Sprout high yield alkali protein bacterial strain bacillus pumilus (the response phase method optimization bacillus pumilus SCU11 of equal breedings divided
Fermentation produce alkali protease and key gene transcriptional control analysis, using with environmental organism journal, 03 phase in 2016) fermentation produce alkali
Property protease activity be 8768U/m L, (bacillus pumilus produces keratinase liquid to one bacillus pumilus of the brave good equal test in Ji
The optimization of fermentation condition, food and fermentation science and technology, 02 phase in 2014) fermenting produces keratinase activity as 57.14U/mL, and Zhang Lei passes through
Screening obtains bacillus pumilus, and (bacillus pumilus TY079 produces the fermentation medium research of antibacterial material, Henan Prov. Inst.
Journal, 02 phase in 2010) there is significant inhibiting effect to cucumber fusarium axysporum, cotton wilt.The screenings such as horse honor obtain short
Bacillus pumilus strain X JAU-117 (screening and Preliminary Identification of Xinjiang walnut rot disease antagonistic bacterium, Xinjiang Agricultural Sciences,
05 phase in 2015), there is wide spectrum antagonistic effect, to the antagonistic effect of walnut rot disease pathogen up to 85%.Therefore, short and small bud
Spore bacillus is a kind of potential biological organic fertilizer fermentation and biological control strain, is found a kind of suitable for the short of tung oil tree grouts
Bacillus pumilus is the direction that those skilled in the art make great efforts research all the time.
Summary of the invention
The technical problem to be solved by the present invention is to overcome the shortcomings of to mention in background above technology and defect, provide one
Bacillus pumilus bacterial strain and its application.
In order to solve the above technical problems, technical solution proposed by the present invention is to provide a bacillus pumilus bacterial strain, it should
Bacillus pumilus (Bacillus pumilus) bacterial strain is preserved in Guangdong Province's Culture Collection (abbreviation GDMCC),
It is named as bacillus pumilus CFC-5 (Bacillus pumilus CFC-5), in Guangdong Province's Microbiological Culture Collection
The deposit number of the heart is GDMCC No:60397, and the preservation time is on June 25th, 2018, and the address of depositary institution is located at Guangzhou
5 building, the building of compound the 59th of martyr Road 100 Guangdong Microbes Inst.
Above-mentioned bacillus pumilus CFC-5 (Bacillus pumilus CFC-5) is isolated and purified from tung oil tree grouts
It obtains, on NA plate, after being inverted 30 DEG C of culture a period of times, colonial morphology is white, flat, transparent poor, microscope
It is in the shape of a rod to observe cell, Gram's staining is the positive.
The DNA gene order of above-mentioned bacillus pumilus CFC-5 (Bacillus pumilus CFC-5), is shown in SEQ ID
Shown in NO.1.
Above-mentioned bacillus pumilus CFC-5 (Bacillus pumilus CFC-5), 16SrDNA sequence exists
That Genbank is compared is bacillus pumilus CFC-5 (Bacillus pumilus CFC-5) and Bacillus as the result is shown
Pumilus (AY876289.1) similitude reaches 100%, therefore is accredited as bacillus pumilus.
Comprehensive morphological observation, Physiology and biochemistry identification and ITS sequence analysis are as a result, the bacillus pumilus can be determined
CFC-5 is bacillus pumilus kind (Bacillus pumilus), is named as bacillus pumilus CFC-5.
Above-mentioned bacillus pumilus CFC-5 (Bacillus pumilus CFC-5), after testing, short and small gemma bar
The noxious material that bacterium CFC-5 (Bacillus pumilus CFC-5) is resistant in tung oil tree grouts and its protein of degrading, it is right
Corps diseases are with wide spectrum inhibiting effect and have growth-promoting functions.
Based on a total technical concept, the present invention correspondingly provides the above-mentioned strain of i (bacillus) pumilus of one kind and is degrading
The application of protein in tung oil tree grouts, application method include the following steps: picking bacillus pumilus CFC-5 to seed culture
Breeding culture (fermenting agent is made) is carried out in base, is then seeded in tung oil tree grouts and is carried out fermented and cultured, it is real in fermentation process
Now to the degradation of protein in tung oil tree grouts.
Above-mentioned application, it is preferred that the seed culture medium includes beef extract, peptone, sodium chloride and deionized water, institute
The mass ratio for stating beef extract, peptone, sodium chloride and water is (3-5): (5-6): (5-6): 1000.
Preferably, controlling pH during the breeding culture is 6.8-7.2, and the temperature for breeding culture is 25 DEG C -45 DEG C,
Time is 12-24h；It is 150-200r/min that revolving speed is controlled in the breeding incubation.
Preferably, the inoculum concentration of the bacillus pumilus CFC-5 is 3-40%, the initial water content of the tung oil tree grouts
For 50%-100%, the temperature of fermented and cultured is 25 DEG C -50 DEG C, and fermentation processes pH is 6.0-8.5, fermentation time 2-8
It.
Preferably, during fermented and cultured, a stirring processing is carried out every 12-48h, it is ensured that bacterial strain can uniformly be sent out
Ferment.
Based on a total technical concept, the present invention correspondingly provides the above-mentioned strain of i (bacillus) pumilus of one kind and is inhibiting
Application in corps diseases.
Above-mentioned application, it is preferred that the plant pathogenic fungi of the corps diseases is phytophthora blight of pepper
(Phytophthora capsici Leonian), flax rhizoctonia solani (Rhizoctonia solaniKuhn), flax anthrax
Germ (Colletotrichum Linicolum Pethybr etLaff), flax wilt bacterium (Fusarium oxysporum
Schl.f.sp.lini (Bolley) Snyder&Hansen), cucumber fusarium axysporum (Fusarium oxysporum
F.sp.cucumebrium Owen) or Sclerotinia sclerotiorum (Sclerotinia scleratiorum (Lib.) deBary).
Compared with prior art, the invention has the benefit that
1, strain of i (bacillus) pumilus provided by the invention is bacillus pumilus CFC-5, which has degradation tung oil tree
The ability of protein in grouts has wide spectrum inhibiting effect to crops soil-borne disease, and has growth-promoting functions, is degraded oil
Paulownia grouts provide bacterium source.
2, bacillus pumilus CFC-5 of the invention can be used as the albumen in fermenting agent fermentative degradation tung oil tree grouts
Matter, tung oil tree cake protein production amount of amino acid of degrading in 100 hours is 127-143mg/g.
3, bacillus pumilus CFC-5 of the invention has inhibiting effect to the growth of plant pathogenic fungi, in phytopathy
There is very big potential using value in harmful prevention and control.
One bacillus pumilus bacterial strain is named as bacillus pumilus CFC-5 (Bacillus pumilus CFC-
5) it, is preserved in Guangdong Province's Culture Collection (abbreviation GDMCC), the guarantor of Culture Collection in Guangdong Province
Hiding number is GDMCC No:60397, and the preservation time is on June 25th, 2018, and the address of depositary institution is located in the martyr of Guangzhou
5 building, the building of compound the 59th of road 100 Guangdong Microbes Inst.
Detailed description of the invention
In order to more clearly explain the embodiment of the invention or the technical proposal in the existing technology, to embodiment or will show below
There is attached drawing needed in technical description to be briefly described, it should be apparent that, the accompanying drawings in the following description is the present invention
Some embodiments for those of ordinary skill in the art without creative efforts, can also basis
These attached drawings obtain other attached drawings.
Fig. 1 is the colonial morphology figure of bacillus pumilus CFC-5 in embodiment.
Fig. 2 is Gram's staining micro-imaging (× 1000) figure of bacillus pumilus CFC-5 in embodiment.
Fig. 3 is the systematic growth tree graph of the 16SrDNA sequence construct of bacillus pumilus CFC-5 in embodiment.
Specific embodiment
To facilitate the understanding of the present invention, the present invention is done below in conjunction with Figure of description and preferred embodiment more complete
Face meticulously describes, but protection scope of the present invention is not limited to following specific embodiments.
Unless otherwise defined, all technical terms used hereinafter are generally understood meaning phase with those skilled in the art
Together.Technical term used herein is intended merely to the purpose of description specific embodiment, and it is of the invention to be not intended to limitation
Protection scope.
Unless otherwise specified, various raw material, reagent, the instrument and equipment etc. used in the present invention can pass through city
Field is commercially available or can be prepared by existing method.
Embodiment:
Bacillus pumilus bacterial strain provided by the invention is named as bacillus pumilus CFC-5 (Bacillus
Pumilus CFC-5), it is preserved in Guangdong Province's Culture Collection (abbreviation GDMCC), in Guangdong Province microorganism fungus kind
The deposit number of collection is GDMCC No:60397, and the preservation time is on June 25th, 2018, and the address of depositary institution is located at
5 building, the building of compound the 59th of Xianlie Middle Road, Guangzhou City 100 Guangdong Microbes Inst.
1, the screening of bacterial strain
1) it tung oil tree grouts source: is sampled from tung oil tree processing enterprise.
2) tung oil tree grouts diffusion juice culture medium: tung oil tree grouts 200g impregnates 3~4h at 60~90 DEG C, with 8 layers of filtered through gauze
It removes residue and obtains paulownia dregs of rice juice, be settled to 1000mL, the packing of 250mL triangular flask, per bottled amount 50mL, pH is natural.
3) tung oil tree grouts fermentation bottom material: tung oil tree grouts and water equal proportion (mass ratio) mix, 500mL conical flask loading amount 40g,
1×105Pa moist heat sterilization 25min (twice).
4) strain primary dcreening operation: tung oil tree grouts and water are mixed in 3: 1 ratios, natural packing fermentation 20d.Take 1g heap fermentation
Tung Oil Cake is inoculated into Tung Oil Cake diffusion juice culture medium, does 3 repetitions, and 30 DEG C, after 180r/min shaking table culture 48h, inhale respectively
Take 1mL bacterial suspension inoculation to Tung Oil Cake diffusion juice culture medium Cao, 30 DEG C, 180r/min shaking table culture 3d.1mL bacterium is taken after 3d respectively
Suspension makees a series of doubling dilution with sterile water.It is respectively 10 by 100 μ L diluted concentrations-13~10-16Dilution be applied to
On NA culture medium, culture is inverted in 30 DEG C of incubators, every thalli growth situation on observation plate for 24 hours.Picking color phenotypes are not
Same bacterium colony carries out scribing line separation, and purifying is repeated, obtains single colonie and is numbered, and 4 DEG C of inclined-planes save.
5) bacterial strain secondary screening: the single strain isolated and purified is inoculated into respectively in 50mLNA fluid nutrient medium, 30 DEG C, 180r/
Min shaking table culture.1mL is sampled when 12h, carries out 10 with sterile water-1~10-8Doubling dilution chooses 10-7~10-8Concentration into
The coating of row NA plate, 3 repetitions of each concentration, 30 DEG C of constant temperature incubations for 24 hours, count the bacterium colony on plate.It is close to bacterium
Degree reaches 108It can be used to inoculation fermentation when CFU/mL.Bacterium solution is inoculated with to tung oil tree grouts fermentation bottom material by 40% inoculum concentration (V/m)
In, 35 DEG C of constant incubator culture 4d, every carrying out turning over sample, the 2 groups of repetitions of each bacterial strain for 24 hours.After fermentation, fermentation material is taken out
70 DEG C of drying, amino-acid nitrogen content, comparison filter out the relatively good bacterial strain of ferment effect in sample before and after Formaldehyde Absorption Method for Determination of Low fermentation.
2, the identification of bacterial strain
1) morphological observation:
Through morphologic observation, as shown in Figure 1, colonial morphology is white, flat, the transparency is poor.As shown in Fig. 2, microscope
Observation presentation cell is in the shape of a rod, and Gram's staining is the positive.
2) Physiology and biochemistry is identified:
Catalase experiment: directly the hydrogen peroxide of dropwise addition 3% is observed immediately, is had in the liquid culture (for 24 hours) of bacterial strain
A large amount of bubble producers are the positive, and not generating bubble, person is feminine gender.Bacterial strain generates a large amount of bubbles immediately, and experimental result is the positive.
Starch Hydrolysis experiment: strain point is connected on starch culture-medium, and a small amount of iodine solution is added dropwise in plate, gently in 30 DEG C of culture 48h
Light rotation, makes iodine solution be uniformly distributed in plate, observes.There is colorless and transparent circle and shows the ability for having hydrolysis starch in periphery of bacterial colonies,
Conversely, not having.Bacterial strain periphery of bacterial colonies is generated without transparent circle, non-hydrolysable starch, for feminine gender.
Methyl red MR experiment: the new pure culture to be tried of picking is a little, is inoculated in collective media, is incubated at 30 DEG C, 3~
5 days, culture solution 1ml is taken, the red indicator 1~2 of methylate drips, and the positive takes on a red color, and weakly positive is in pale red, and feminine gender is yellow.Bacterium
Liquid reddens color, for the positive.
VP experiment: the new pure culture of picking is a little, is inoculated in collective media, first in 30 DEG C of culture 2d, culture solution 2.5ml
A- naphthols absolute alcohol solution 0.6ml, then plus 40% potassium hydroxide aqueous solution 0.2ml, shake 2~5min, positive bacteria Chang Li is added
Red is presented, if redfree occurs, being statically placed in room temperature or 30 DEG C of insulating boxs be can determine that if do not shown red still in 2h as yin
Property.Bacterium solution reddens immediately, for the positive.
Gelatin liquefaction experiment: new pure culture is taken, percutaneous puncture-inoculation separately there are two not to be inoculated in about 2/3 depth of gelatin high level
Blank control.3~5d is cultivated at 20 DEG C.Observation daily is as a result, if bacterial growth and gelatin partly or entirely becomes flowable
Liquid, then be that test is positive.It otherwise is feminine gender.Gelatin is liquefied, for feminine gender.
Nitrate reduction experiment: culture is inoculated in nitrate broth culture medium, 28 DEG C of shaking table culture 3d take 5mL to train
Nutrient solution illustrates that color developing agent is added by kit (Hai Bo Bioisystech Co., Ltd nitrate reduction kit), turned yellow as the positive,
Feminine gender is not changed color as.Bacterium solution discoloration, for the positive.
Produce hydrogen sulfide experiment: by bacterium percutaneous puncture-inoculation to be checked in lead acetate medium, in 35 DEG C of 24~48h of culture observation knots
Fruit.Culture medium blackening is the positive, does not become negative.Culture medium does not change colour, for feminine gender.
Cellulose decomposition experiment: by strain to be tested streak inoculation in the agar based media for adding 0.8% cellulose powder
On, using the culture medium that is not inoculated with as blank control, 30 DEG C are cultivated 1~2 week, and bacterial strain can be grown on culture medium, experimental result
For the positive；It otherwise is feminine gender.Bacterial strain is not grown, for feminine gender.
Phosphorus decomposing test: by the NBRIP culture medium of strain inoculated to 2% agar of addition, 30 DEG C of culture 3d, periphery of bacterial colonies has
It is colorless and transparent to iris out existing person as the positive, it was demonstrated that the bacterial strain has dissolving P capacity.Bacterial strain periphery of bacterial colonies has colourless transparent circle to occur, and is
It is positive.
It produces heteroauxin (IAA) test: strain to be tested is inoculated into the TSB culture medium of addition 0.5mmol/L L-Trp
In 30 DEG C of shaking table culture 48h, take culture solution 2mL, 10000r/min is centrifuged 10min, takes supernatant, and every 1mL supernatant adds 2mL
Salkowski reagent, room temperature dark place stand 30min, and using blank cultures as control, generating then explanation if any pink colour has IAA
It generates.There is pink colour generation, have and produce IAA ability, for the positive.
Produce ammonia test: by strain inoculated into peptone culture medium, 30 DEG C of shaking table culture 3d take culture solution a little in test tube
In, 5 drop nessler reagents are added dropwise, using blank cultures as control, tan precipitate occur for the positive, it was demonstrated that bacterial strain, which has, produces ammonia
Ability.There is tan precipitate generation, have and produce ammonia ability, for the positive.
To sum up, Physiology and biochemistry qualification result such as table 1.
Table 1: the Physiology and biochemistry qualification result of this bacterial strain
| Table feature | Response feature |
| Contact enzymatic determination | + |
| Starch Hydrolysis measurement | - |
| Methyl red MR measurement | + |
| VP experiment | + |
| Gelatin liquefaction measurement | - |
| Nitrate reduction measurement | + |
| Produce hydrogen sulfide measurement | - |
| Cellulose decomposition | - |
| Phosphorus decomposing test | + |
| Produce IAA test | + |
| Produce ammonia test | + |
Wherein, "+" be expressed as this bacterial strain have reaction or can use, "-" be shown as this bacterial strain do not react or cannot benefit
With.
1 test result of table illustrates that bacterial strain CFC-5 can produce heteroauxin, has dissolving P capacity, produces ammonia ability, these
Growth-promoting attribute testing result illustrates that bacterial strain CFC-5 has the potentiality for promoting plant growth.
3) the 16SrDNA sequencing of bacterial strain:
Candidate strain DNA is extracted using DNA of bacteria extracts kit, is expanded using bacterium amplification universal primer 16SF and 16SR
Increase the 16S rRNA of bacterium；PCR reaction system (50 μ L) is 10 × PCRbuffer, 5 μ L, DNA profiling 1 μ L, dNTP
(2.5mmol/L) 4 μ L, MgCl2 (25mmol/L) 3 μ L, each 0.5 μ L of 1 μ L, Taq archaeal dna polymerase of primer (1mmol/L), adds
ddH2O to 50 μ L；Reaction condition is 94 DEG C of 5min, 94 DEG C of 30s, 53 DEG C of 30s, 72 DEG C of 1min, and totally 35 recycle, 4 DEG C of preservations.Expand
Volume increase object be sequenced by Shanghai Sheng Gong bioengineering Co., Ltd, sequencing result as shown in sequence table SEQ ID NO.1,
The systematic growth tree graph of 16SrDNA sequence construct is as shown in Figure 3.
3. application of the bacillus pumilus CFC-5 to corps diseases inhibiting effect
As shown in table 2, bacterial strain CFC-5 is vertical to phytophthora blight of pepper (Phytophthora capsici Leonian), flax
Blight bacterium (Rhizoctonia solaniKuhn), anthracnose of flax bacterium (Colletotrichum Linicolum Pethybr
EtLaff), flax wilt bacterium (Fusarium oxysporum Schl.f.sp.lini (Bolley) Snyder&Hansen),
Cucumber fusarium axysporum (Fusarium oxysporum f.sp.cucumebrium Owen), Sclerotinia sclerotiorum
The growth of 6 plant pathogenic fungis such as (Sclerotinia scleratiorum (Lib.) deBary) has inhibiting effect, right
Oil Tea Anthracnose bacterium rejection ability is most strong, antibacterial circle diameter 23.29mm；It is weaker to anthracnose of flax bacterium rejection ability, but press down
Bacterium loop diameter has also reached 17.7mm.Illustrate that bacterial strain CFC-5 is the Antagonistic Fungi of one plant of high-efficiency broad spectrum, in the prevention and control of plant disease
With very big potential using value.
Table 2: bacteriostasis of the bacterial strain CFC-5 to 6 plant pathogenic fungis
| Disease fungus | Antibacterial circle diameter (mm)a |
| P. capsici | 19.28±1.20bc |
| Flax rhizoctonia solani | 22.00±0.48ab |
| Anthracnose of flax bacterium | 17.70±0.43e |
| Oil Tea Anthracnose bacterium | 23.29±0.47a |
| Cucumber fusarium axysporum | 20.00±0.23cd |
| Sclerotinia sclerotiorum | 19.71±0.84d |
Data are average value ± standard error in table, significant difference (P < 0.05) between different lowercase expression processing.
4, application of the bacillus pumilus CFC-5 as albumen in leavening and degradation tung oil tree grouts
1) selection of culture medium
Seed culture medium: beef extract 3g, peptone 5g, sodium chloride 5g, deionized water 1000mL adjust pH to 6.8-7.2,
50mL is dispensed into 250mL conical flask, 121 DEG C of sterilizing 20min.
Fermentation medium: tung oil tree grouts (40 mesh) 100g, water 25mL is loaded in 1000mL conical flask, pH 6.8-7.2,
121 DEG C of sterilizing 20min.
2) implementation method
Bacillus pumilus CFC-5 on one ring slant medium of picking, until in seed culture medium, 35 DEG C of cultivation temperature,
Incubation time is 20h, revolving speed 180r/min；It is seeded in fermentation medium with 15% inoculum concentration later, incubation time is
6d, cultivation temperature are 45 DEG C, during which stir one time fermentation bottle every 12h.The crude protein measured in fermentation flask after fermentation contains
Amount and amino nitrogen content.
3) detection method
Biomass estimation method is counted using dilution plating procedure.
The measurement of amino nitrogen uses formaldehyde method: accurately weighing tung oil tree grouts 5g in 250mL beaker, 40mL distillation is added
Water.2-3min is boiled in electric furnace heating.It is cooled to room temperature, supernatant transfer is settled to 50mL volumetric flask, draws 10mL supernatant, makes
Formalin 10mL is added after being adjusted to 8.20 with pH meter, being titrated to 9.20 with 0.5mol/L standard solution of sodium hydroxide is terminal,
It writes down consumption sodium hydroxide volume and calculates amino-acid nitrogen content.Reagent blank test is done simultaneously, calculation method is as follows:
In formula:
Y-amino-acid nitrogen content (in terms of nitrogen)/g；
V0- sample spends standard solution of sodium hydroxide number/mL after formaldehyde is added；
Handy standard solution of sodium hydroxide number/mL after formaldehyde is added in V-reagent blank；
Vl- draw dilution number/mL；
The concentration of N-NaOH standard solution/(mol/L)；
Milliequivalent/g of the suitable nitrogen of 0.014-consumption 1mLNaOH standard solution.
4) testing result
| Index | Before fermentation | After fermentation |
| Amino nitrogen | 16.52mg/g | 420.81mg/g |
| Bacillus pumilus biomass | 0 | 2.1×1019CFU/g |
Can significantly it find out from testing result, amino nitrogen and bacillus pumilus biomass have bright after fermentation
Aobvious raising, the amount of amino nitrogen is increased to 421mg/g by the 16.5mg/g before fermenting after fermentation 6 days, amino nitrogen content compared with
Before increase 25.47 times, bacillus pumilus biomass has also reached 2.1 × 1019CFU/g illustrates the strain in tung oil tree grouts
Growth fermentation is all right, the effect of protein (being indicated with the amount that protein breakdown is amino nitrogen) in tung oil tree grouts of degrading
It is good, and the noxious material being resistant in tung oil tree grouts.
Embodiment described above is merely a preferred embodiment of the present invention, and the simultaneously exhaustion of the feasible implementation of non-present invention.It is right
For persons skilled in the art, any aobvious to made by it under the premise of without departing substantially from the principle of the invention and spirit and
The change being clear to should be all contemplated as falling within claims of the invention.
Sequence table
<110>Sino-South African Forestry University of Science and Technology
<120>one bacillus pumilus bacterial strains and its application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1450
<212> DNA
<213>bacillus pumilus CFC-5 (Bacillus pumilus CFC-5)
<400> 1
gggcaggcgg ctgctataca tgcagtcgag cggacagaag ggagcttgct cccggatgtt 60
agcggcggac gggtgagtaa cacgtgggta acctgcctgt aagactggga taactccggg 120
aaaccggagc taataccgga tagttccttg aaccgcatgg ttcaaggatg aaagacggtt 180
tcggctgtca cttacagatg gacccgcggc gcattagcta gttggtgagg taacggctca 240
ccaaggcgac gatgcgtagc cgacctgaga gggtgatcgg ccacactggg actgagacac 300
ggcccagact cctacgggag gcagcagtag ggaatcttcc gcaatggacg aaagtctgac 360
ggagcaacgc cgcgtgagtg atgaaggttt tcggatcgta aagctctgtt gttagggaag 420
aacaagtgca agagtaactg cttgcacctt gacggtacct aaccagaaag ccacggctaa 480
ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggaa ttattgggcg 540
taaagggctc gcaggcggtt tcttaagtct gatgtgaaag cccccggctc aaccggggag 600
ggtcattgga aactgggaaa cttgagtgca gaagaggaga gtggaattcc acgtgtagcg 660
gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag gcgactctct ggtctgtaac 720
tgacgctgag gagcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780
cgtaaacgat gagtgctaag tgttaggggg tttccgcccc ttagtgctgc agctaacgca 840
ttaagcactc cgcctgggga gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg 900
cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt 960
cttgacatcc tctgacaacc ctagagatag ggctttccct tcggggacag agtgacaggt 1020
ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc 1080
aacccttgat cttagttgcc agcattcagt tgggcactct aaggtgactg ccggtgacaa 1140
accggaggaa ggtggggatg acgtcaaatc atcatgcccc ttatgacctg ggctacacac 1200
gtgctacaat ggacagaaca aagggctgcg agaccgcaag gtttagccaa tcccacaaat 1260
ctgttctcag ttcggatcgc agtctgcaac tcgactgcgt gaagctggaa tcgctagtaa 1320
tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca 1380
ccacgagagt ttgcaacacc cgaagtcggt gaggtaacct ttatggagcc agccgccgaa 1440
gtggcaagtt 1450
Claims (9)
1. a bacillus pumilus bacterial strain, which is characterized in that bacillus pumilus (Bacilluspumilus) the bacterial strain preservation
In Guangdong Province's Culture Collection, it is named as bacillus pumilus CFC-5 (Bacilluspumilus CFC-5),
It is GDMCC No:60397 in the deposit number of Guangdong Province's Culture Collection, and the preservation time is June 25 in 2018
Day.
2. a kind of application of strain of i (bacillus) pumilus according to claim 1 protein in degradation tung oil tree grouts.
3. application according to claim 2, which is characterized in that its application method includes the following steps: the short and small gemma of picking
Bacillus CFC-5 carries out breeding culture into seed culture medium, is then seeded in the fermentation medium containing tung oil tree grouts and carries out
Fermented and cultured realizes the degradation to protein in tung oil tree grouts in fermentation process.
4. application according to claim 3, which is characterized in that the seed culture medium includes beef extract, peptone, chlorination
Sodium and deionized water, the beef extract, peptone, sodium chloride and water mass ratio be (3-5): (5-6): (5-6): 1000.
5. application according to claim 3, which is characterized in that controlling pH during the breeding culture is 6.8-7.2,
The temperature of breeding culture is 25 DEG C -45 DEG C, time 12-24h；It is 150-200r/ that revolving speed is controlled in the breeding incubation
min。
6. application according to claim 3, which is characterized in that the inoculum concentration of the bacillus pumilus CFC-5 is 3-
40%, the initial water content of the tung oil tree grouts is 50%-100%, and the temperature of fermented and cultured is 25 DEG C -50 DEG C, fermentation process
Control pH is 6.0-8.5, and fermentation time is 2-8 days.
7. the application according to any one of claim 3-6, which is characterized in that during fermented and cultured, every 12-48h
Carry out a stirring processing, it is ensured that bacterial strain being capable of uniform fermentation.
8. a kind of strain of i (bacillus) pumilus according to claim 1 is inhibiting the application in corps diseases.
9. application according to claim 8, which is characterized in that the plant pathogenic fungi of the corps diseases is capsicum epidemic disease
Mould (Phytophthora capsiciLeonian), flax rhizoctonia solani (Rhizoctonia solaniKuhn), flax charcoal
Subcutaneous ulcer germ (Colletotrichum Linicolum Pethybr etLaff), flax wilt bacterium (Fusarium
Oxysporum Schl.f.sp.lini (Bolley) Snyder&Hansen), cucumber fusarium axysporum (Fusarium
Oxysporum f.sp.cucumebrium Owen) or Sclerotinia sclerotiorum (Sclerotinia scleratiorum (Lib.)
deBary)。
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| CN111826327B (en) * | 2020-08-06 | 2023-05-02 | 清远一生自然生物研究院有限公司 | Bacillus pumilus BP-09 capable of tolerating high-concentration tea residues and application thereof |
| CN112608874A (en) * | 2021-01-19 | 2021-04-06 | 中国科学院兰州化学物理研究所 | Bacillus bacteria with growth promoting activity and preparation method and application thereof |
| CN113215043A (en) * | 2021-05-12 | 2021-08-06 | 宁波浮田生物技术有限公司 | Bacillus pumilus and application thereof |
| CN116064282A (en) * | 2022-07-29 | 2023-05-05 | 贵州大学 | Fermentation technology for rapidly degrading tea saponin of cold pressed cake of oil tea and special fungus thereof |
| CN116064282B (en) * | 2022-07-29 | 2024-04-05 | 贵州大学 | Fermentation technology for degrading tea saponin of cold pressed cake of oil tea and special fungus thereof |
| CN115812744A (en) * | 2022-10-28 | 2023-03-21 | 中南林业科技大学 | Tung tree extract and application thereof in inhibiting anthracnose of camellia oleifera |
| CN115812744B (en) * | 2022-10-28 | 2023-11-21 | 中南林业科技大学 | Tung tree extract and application thereof in inhibiting anthracnose of oil tea |
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