CN105907680A - Bacillus subtilis J-5 as well as biological agent and application thereof - Google Patents

Bacillus subtilis J-5 as well as biological agent and application thereof Download PDF

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CN105907680A
CN105907680A CN201610359661.2A CN201610359661A CN105907680A CN 105907680 A CN105907680 A CN 105907680A CN 201610359661 A CN201610359661 A CN 201610359661A CN 105907680 A CN105907680 A CN 105907680A
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bacillus subtilis
fructus
strain
spore
application
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CN201610359661.2A
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CN105907680B (en
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贾振华
金伟伟
黄亚丽
宋水山
马宏
宋聪
张立文
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河北省科学院生物研究所
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12RPROCESSES USING MICROORGANISMS
    • C12R1/00Processes using microorganisms
    • C12R1/01Processes using microorganisms using bacteria or actinomycetales
    • C12R1/07Bacillus
    • C12R1/125Bacillus subtilis
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01NPRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/10Animals; Substances produced thereby or obtained therefrom

Abstract

The invention provides bacillus subtilis having a preservation number of CGMCC No.11750, a microbial agent containing the bacillus subtilis and application of the bacillus subtilis or microbial agent in prevention and control of grey mold of tomatoes, cucumbers, strawberries and other fruits and vegetables. The bacillus subtilis having the preservation number of CGMCC No.11750 and the microbial agent containing the bacillus subtilis have a strong antagonizing effect on botrytis cinerea, have quick response and have long prevention lasting period. A corresponding microbial preparation can be prepared by utilizing the screened bacillus subtilis strains, and occurrence of grey mold on tomatoes, cucumbers, strawberries and other vegetables can be prevented and controlled by the bacillus subtilis after the prepared microbial agent is applied to the field, so that chemical pesticide application in the fruit and vegetable production process can be reduced.

Description

A kind of bacillus subtilis J-5 and bacteria agent thereof and application
Technical field
The invention belongs to biological technical field, be specifically related to a kind of bacillus subtilis J-5 and bacteria agent thereof and application.
Background technology
Botrytis cinerea (Botrytis cinerea), also known as the pathogen of Botrytis cinerea, is a universal fungus of class, and it is one Downright bad auxotype cause of disease that plant wide host's property, that more than 200 kind of known plants (such as water fruits and vegetables and flowers) gray mold can be caused Fungus.Survive the winter with invalid being organized in soil with sclerotium, conidium and mycelium, by wound, natural aperture and young tender group Knit intrusion host, a large amount of conidium can be produced again after morbidity, infect the most again by means of wind and rain its spread in china, cause grey mold Disease, flower, really, leaf, stem all can fall ill, cause crop to wilt death, affect yield and quality, be to produce the weight that upper preventing and treating is the most difficult Want one of disease.
On Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, Fructus Fragariae Ananssae etc., gray mold disease Seedling color is shallow, and blade, petiole are fallen ill in canescence, water Stain shape, organization softening is to rotting, and during high humidity, there is grey mold on surface.Children's stem is many comes into being irregular water logging speckle at petiole base, quickly Deliquescing is rotted, and hangs and contracts or roll over, and last sick scutellum rot is withered and died of illness.Fruit is caught an illness, and Fructus Canarii is injured heavily, the stigma of residual or petal The most first being infected, backward fruit extends, and causing peel is canescence, and has the mould layer of thick Lycoperdon polymorphum Vitt, and in water corruption shape, blade is sent out Sick from the beginning of blade tip, become " V " shape inwardly extended along between vein, taupe, while there is the alternate stricture of vagina shape line of the depth, sick key has a common boundary point Bright.Catch an illness the light then underproduction 20%, the serious underproduction more than 50% in plot.
At present gray mold has become a kind of Major Diseases on Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, strawberry production, to Fructus Cucumidis sativi, kind Eggplant, eggplant, Fructus Capsici, Yield of Strawberry and quality constitute the most serious threat, and this disease is currently mainly by antibacterial class chemistry agriculture Medicine is prevented and treated, but poor effect and environmental pollution are more serious, is badly in need of finding the gray mold preventing and treating of low stain, high preventive effect Mode.
Utilize antagonistic microbe preventing and treating corps diseases owing to having low stain, low-residual, not causing Disease Resistance etc. excellent Point, is paid attention to the most widely.This controlling mode it is crucial that filter out and pathogenic microorganism had antagonism and antibacterial Compose wide antagonistic microbe, and utilize microbial inoculum or its metabolite crude extract to form biological preparation, Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, the gray mold of Fructus Fragariae Ananssae are prevented and treated and are applied.The most some are for biological prevention and control agents of Biological Control of Gray Mold, but greatly Many fungistatic effects are not ideal enough, and antimicrobial spectrum is narrow, and acting duration is short.Therefore, in the urgent need to filter out one can efficiently, The microbial strains of lasting antagonism gray mold source of disease bacterium and relevant microbial bacterial agent, thus reduce gray mold by Biological control The harm brought to Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, strawberry production.
Summary of the invention
Based on the above state of the art, it is an object of the invention to provide one effectively, persistently can suppress botrytis cinerea to grow, Especially can the bacillus subtilis strain of efficiently antagonism Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, grey mould fruit rot of strawberry, and the present invention is also Provide a kind of containing above-mentioned bacillus subtilis strain for prevent and treat botrytis cinerea infect caused by Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, sweet Green pepper, the bacteria agent of grey mould fruit rot of strawberry.
In order to reach foregoing invention purpose, the technical solution adopted in the present invention is as follows:
Bacillus subtilis J-5 of the present invention on November 27th, 2015 in China Committee for Culture Collection of Microorganisms General Microbiological Culture preservation center has carried out preservation, and preserving number is: CGMCC No. 11750.
Present invention also offers the bacteria agent containing described bacillus subtilis J-5, wherein bacillus subtilis J-5 Content is 4 × 109cfu/g~7×109Cfu/g microbial inoculum.
Described bacteria agent is the mix bacterium agent of the spore microbial inoculum of bacillus subtilis J-5.
Present invention also offers described bacillus subtilis J-5 and microbial inoculum thereof preventing and treating Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, Fructus Fragariae Ananssae Deng the application in fruit and vegerable gray mold.
The invention have the benefit that
The bacillus subtilis J-5 of the present invention can effectively suppress the Fructus Cucumidis sativi caused by botrytis cinerea infection, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, Fructus Fragariae Ananssae Gray mold, has efficient preventive and therapeutic effect, and effective percentage can reach more than 60%, and instant effect, and acting duration is long.As Biological intervention means, can avoid residual and pollution that chemicals such as pesticide causes, are a kind of more economical, environmental protection, the planting of green Thing disease control means.
The bacillus subtilis J-5 of the present invention can make multiple bacteria agent, or add to existing as effective ingredient In bacteria agent, when making spore microbial inoculum, it is suitable for multiple Conservation environment, and there is the longer shelf-life, at normal temperature and pressure Under, 1-3 can be preserved, be beneficial to storage and transport.
Biological sample preservation information
Bacillus subtilis J-5, Classification And Nomenclature isBacillus subtilis, it is preserved in China on November 27th, 2015 Microbiological Culture Collection administration committee common micro-organisms center, is called for short CGMCC, address: BeiChen West Road, Chaoyang District, BeiJing City 1 Institute 3, Institute of Microorganism, Academia Sinica, postcode 100101, culture presevation number is CGMCC No. 11750.
Accompanying drawing explanation
Fig. 1 is the colonial morphology that bacillus subtilis J-5 cultivates 48 hours on solid LB media.
Fig. 2 is the form under the optical microscope after the dyeing of bacillus subtilis J-5 azaleine.
Detailed description of the invention
Embodiment 1 has the separation of antibacterial of antagonism, screening to Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, grey mould fruit rot of strawberry
Gather the healthy plant rhizosphere soil in the morbidity of grey mold disease serious Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, Fructus Fragariae Ananssae plot as examination Test material, in laboratory, from this sample, carry out bacteria distribution, LB culture medium carries out separating for several times purification, obtains pure training Bacteria falls behind, and with botrytis cinerea as indicator bacteria, uses mutual confrontation test method to carry out the screening of the high bacterial strain of antagonism performance.
Concrete steps: above-mentioned pedotheque from Hebei province, Ningxia, Shandong, the Huang being infected with grey mold disease on the ground such as Shanxi In melon, Fructus Lycopersici esculenti, eggplant, Fructus Capsici, strawberry cultivating field, this soil is coated after gradient dilution in PB culture medium, in 28 DEG C of perseverances Temperature incubator is cultivated.Above-mentioned culture medium prepares according to the record of " microbiological culture media handbook ", and culture medium prescription is: beef Cream 5g, peptone 10g, sodium chloride 5g, agar 15g, tap water 1000ml, pH7.2 ~ 7.4, use after 121 DEG C of sterilizing 20min. After flat board grows bacterium colony, single bacterium colony of picking different shape uses line partition method to be repeated isolated and purified, until obtaining The pure culture of single bacterium colony, bacterial strain after purification is stored in PB medium slant.
With tomato gray mould bacterium as indicator bacteria, decussation method is used to measure the suppression ratio that botrytis cinerea is grown by different strains. The bacterial isolates filtered out point is connected on 4 angles at anomaly plate central authorities 3cm, flat board central authorities inoculation diameter 5mm's simultaneously Activation graw mold of tomato truffle, each bacterial strain in triplicate, 28 DEG C of constant temperature culture 5d, measure, record the big of antibacterial circle diameter Little and calculate average inhibition.Filter out the bacterial strain J-5 the strongest to tomato gray mould bacterium antagonism.
And this bacterial strain has been carried out preservation, preserving number is CGMCC No.11750.
The determination of embodiment 2 antagonistic strain hereditary stability
Bacterial strain J-5 uses in PB culture medium method of scoring carry out Secondary Culture, after cultivating for 20 generations, uses flat board face-off method to measure The J-5 bacterial strain after the passing on antagonistic effect to tomato gray mould bacterium, with original J-5 bacterial strain for comparison, this antagonistic strain is through 20 generations There is with original strain after cultivation identical antagonism, illustrate that this antagonistic strain has good hereditary stability, by antibacterial The Antagonistic Fungi of effect stability carries out PB medium liquid cultivation, draws bacterium solution and carries out-80 DEG C of glycerol preservations.
The qualification of embodiment 3 antagonistic strain
According to the experiment content described in " common bacteria system identification handbook " and experimental technique, to the preserving number sieved it is The bacterial strain J-5 of CGMCC No.11750 identifies, colonial morphology observation is carried out in PB culture medium, bacterial strain line is connect After Zhong, within 24 hours, observe colonial morphology in 28 DEG C of constant temperature culture.Preserving number is that the bacterium colony of the bacterial strain of CGMCC No.11750 is round Shape, edge erodes shape, umbilical hernia shape projection, surface ruffle, and color is milky, matt opaque, is combined the tightest with culture medium Close.At 100 times of oily Microscopic observations of optical microscope after picking 24 hours lawn Gram’s staining of cultivation.Under the microscope may be used To see, preserving number be the bacterial strain of CGMCC No.11750 be bacillus cereus, gram positive bacteria, in spore position raw or near in Raw, spore expands, without parasporal crystal, and single appearance.Primarily determine that as bacillus cereus.
16SrDNA identifies: the inoculation that preserving number is CGMCC No.11750 trained to PB culture fluid, 28 DEG C of shaking tables Support 24 h, use TIANGEN Biotech's genome DNA extracting reagent kit to extract the genome of this bacterial strain DNA, then using extract STb gene as template, with forward primer: F27(5 '-AGAGTTTGATCCTGGCTCAG-3 ') and under Trip primer R1492(5 '-TACGGCTA CCTTGTTA CGAC TT-3 ') primer is to carrying out PCR amplification.PCR reaction system Being 25 μ l, reaction system is: genomic DNA 1 μ L, 10 × PCR buffer 2.5 μ L, F27Primer 0.5 μ L, R1492Primer 0.5 μ L, dNTPs 2 μ L, Taq enzyme (5U/ μ L) 0.25 μ L, ddH2O 18.25 μ L.Reaction condition is: 95 DEG C of denaturation 5 min; 95 DEG C of degeneration 30s, 50 DEG C of annealing 1min, 72 DEG C extend 2min, circulate 30 times;72℃ 10 min.PCR primer 1% agarose Gel carries out electrophoresis detection, the PCR primer of PCR stripe size about 1500bp is served Hai Sheng work Bioisystech Co., Ltd and surveys After sequence, the DNA sequence (see sequence 1) that will obtain, input GenBank, carry out with all sequences in data base by Blast program Comparative analysis.MEGA4.1 is utilized to carry out the structure of phylogenetic tree.Showing through sequence analysis, preserving number is CGMCC The bacterial strain of No.11750 belongs to bacillus subtilis.
The tomato planting Field information of embodiment 4-7 bacillus subtilis J-5
It is that CGMCC No.11750 bacillus subtilis strain is inoculated into the 30L equipped with 15L PB fluid medium and sends out by preserving number In ferment tank, in 28 DEG C, cultivate 36 hours under the constant temperature of 180r/min, the centrifugal spore obtaining bacillus subtilis, then It is spray-dried, mixes the spore dry powder of this bacterium with soluble starch, form final concentration of 4 × 109cfu/g~7×109Cfu/g's Cultivated spore preparation.
Fructus Lycopersici esculenti continuous cropping field in Qing Country, Hebei Province and Botou City two area carries out the field of bacillus cereus agent to graw mold of tomato Between test.The weight ratio of cultivated spore preparation and water 1:300 is hanged molten after spray respectively after Fructus Lycopersici esculenti field planting 10 days, florescence and first fruit Phase, each using dosage is every mu of 300g cultivated spore preparation;Routine control uses the high skilful conduct preventing and treating of chemical pesticide of routine dose Medicine, blank is not used protective agents but is applied the water with embodiment group identical weight;Each process repeats for 3 times, investigation The incidence of graw mold of tomato.
Investigation method:
Using diagonal five point sampling in the period of maturation, each point determines strain 20 strain, samples 100 strain investigation strain sickness rate and the state of an illness altogether Index, record total investigation strain number, total morbidity strain number.
Graw mold of tomato disease index grade scale:
0 grade: the harmless scab of Herb;1 grade: the blade of minority blade morbidity to Herb bottom less than 1/4 has scab;2 grades: Herb The blade of middle and lower part less than 1/2 has on scab or stem stalk branch and has scab;3 grades: Herb 3/4 has scab with the blade of lower portion Or single branch stem stalk is withered and individual plant has 1 ~ 2 fruit morbidity to rot;4 grades: the morbidity of the blade of Herb more than 3/4 withered and yellow or Person's stem stalk is withered, and individual plant has the fruit morbidity of more than 2 to rot.
Preventive effect computational methods:
Tomato strain sickness rate=total morbidity strain number/total investigation strain number × 100
Disease index=∑ (morbidity strain number × this morbidity strain level value)/(total investigation strain number × superlative degree value) × 100
Prevention effect (%)=(check plot disease index treatment region disease index)/disease index × 100, check plot
Prevention effect is as shown in table 1, and J-5 spore microbial inoculum has good preventive effect to graw mold of tomato, is using J-5 spore microbial inoculum In the case of graw mold of tomato sickness rate and disease index all reduce, the protection effect of different embodiments is all more than 80%.
The prevention effect to graw mold of tomato used by the different microbial inoculum of table 1
The green cucumber Field information of embodiment 8-11 bacillus subtilis J-5
It is that CGMCC No.11750 bacillus subtilis strain is inoculated into the 30L equipped with 15L PB fluid medium and sends out by preserving number In ferment tank, in 28 DEG C, cultivate 36 hours under the constant temperature of 180r/min, the centrifugal spore obtaining bacillus subtilis, then It is spray-dried, mixes the spore dry powder of this bacterium with soluble starch, form final concentration of 4 × 109cfu/g~7×109Cfu/g's Cultivated spore preparation.
In the Fructus Cucumidis sativi field in Hebei province's Hengshui City and Botou City two area, gray mold is carried out the field examination of bacillus cereus agent Test.The weight ratio of cultivated spore preparation and water 1:300 is hanged molten after spray respectively after Fructus Cucumidis sativi field planting 10 days and florescence, use every time Dosage is every mu of 300g cultivated spore preparation;Routine control uses the chemical pesticide height of routine dose ingeniously as protective agents, and blank is right The water with embodiment group identical weight is applied according to not using protective agents;Each process repeats for 3 times, investigation gray mold of cucumber Incidence.
Investigation method:
Using diagonal five point sampling in the period of maturation, each point determines strain 20 strain, samples 100 strain investigation strain sickness rate and the state of an illness altogether Index, record total investigation strain number, total morbidity strain number.
Gray mold of cucumber disease index grade scale:
0 grade: without disease;
1 grade: a small amount of scab occurs in inoculation leaf;
2 grades: lesion area accounts for the 1/3 of leaf area;
3 grades: lesion area accounts for the 1/3-1/2 of leaf area;
4 grades: lesion area accounts for the 1/2-2/3 of leaf area;
5 grades: lesion area accounts for more than the 2/3 of leaf area
Preventive effect computational methods:
Fructus Cucumidis sativi strain sickness rate=total morbidity strain number/total investigation strain number × 100
Disease index=∑ (morbidity strain number × this morbidity strain level value)/(total investigation strain number × superlative degree value) × 100
Prevention effect (%)=(check plot disease index treatment region disease index)/disease index × 100, check plot
J-5 spore microbial inoculum is utilized to carry out the controlling experiment of gray mold of cucumber, result in Hebei province's Hengshui City and Botou City two area Being shown in Table 2, use this spore microbial inoculum can reduce gray mold of cucumber sickness rate, reduce disease index, protection effect is more than 80%.
The prevention effect to gray mold of cucumber used by the different microbial inoculum of table 2
The strawberry cultivating Field information of embodiment 12-15 bacillus subtilis J-5
It is that CGMCC No.11750 bacillus subtilis strain is inoculated into the 30L equipped with 15L PB fluid medium and sends out by preserving number In ferment tank, in 28 DEG C, cultivate 36 hours under the constant temperature of 180r/min, the centrifugal spore obtaining bacillus subtilis, then It is spray-dried, mixes the spore dry powder of this bacterium with soluble starch, form final concentration of 4 × 109cfu/g~7×109Cfu/g's Cultivated spore preparation.
Strawberry continuous cropping booth in Luancheng, Hebei province and area, Handan two carries out the field examination of bacillus cereus agent to gray mold Test.The weight ratio of cultivated spore preparation and water 1:300 is hanged molten after spray at the beginning of Fructus Fragariae Ananssae fruit and plant leaf on, cross within seven days, investigate Gray mold situation, using dosage is every mu of 100 grams of cultivated spore preparations;Routine control uses the high skilful conduct of chemical pesticide of routine dose Protective agents, blank is not used protective agents but is applied the water with embodiment group identical weight;Each process repeats for 3 times, The incidence of investigation grey mould fruit rot of strawberry and yield.
Investigation method:
Using diagonal five point sampling in the period of maturation, each point determines strain 20 strain, samples 100 strain investigation strain sickness rate and the state of an illness altogether Index, record total investigation strain number, total morbidity strain number.
Grey mould fruit rot of strawberry disease index grade scale:
0 grade: without disease;
1 grade: lesion area accounts for less than the 5% of whole fruit face;
3 grades: lesion area accounts for less than the 6% ~ 10% of whole fruit face;
5 grades: lesion area accounts for less than the 11% ~ 25% of whole fruit face;
7 grades: lesion area accounts for less than the 26% ~ 50% of whole fruit face;
9 grades: lesion area accounts for more than the 50% of whole fruit face.
Preventive effect computational methods:
Tomato strain sickness rate=total morbidity strain number/total investigation strain number × 100
Disease index=∑ (morbidity strain number × this morbidity strain level value)/(total investigation strain number × superlative degree value) × 100
Prevention effect (%)=(check plot disease index treatment region disease index)/disease index × 100, check plot
Prevention effect is as shown in table 3, uses this spore microbial inoculum can reduce grey mould fruit rot of strawberry sickness rate, reduce disease index, within 7 days, prevents Sick effect is i.e. up to more than 90%.
The prevention effect to grey mould fruit rot of strawberry used by the different microbial inoculum of table 3
By the above results it can be seen that the bacterial strain that preserving number is CGMCC No.11750 that filters out of the present invention is to Fructus Fragariae Ananssae ash Mildew has good antagonism, and the generation of strawberry greenhouse gray mold can effectively be prevented and treated by its spore microbial inoculum.
The Fructus Capsici plantation Field information of embodiment 16-19 bacillus subtilis J-5
It is that CGMCC No.11750 bacillus subtilis strain is inoculated into the 30L equipped with 15L PB fluid medium and sends out by preserving number In ferment tank, in 28 DEG C, cultivate 36 hours under the constant temperature of 180r/min, the centrifugal spore obtaining bacillus subtilis, then It is spray-dried, mixes the spore dry powder of this bacterium with soluble starch, form final concentration of 4 × 109cfu/g~7×109Cfu/g's Cultivated spore preparation.
(disease index is to have infected the Sweet pepper seedlings of gray mold in Luancheng, Hebei province and area, Handan two Fructus Capsici continuous cropping booth 1.25%) field test of bacillus cereus agent is carried out.The weight ratio of cultivated spore preparation and water 1:300 is hanged molten after spray at Fructus Capsici leaf On sheet, mu consumption 100g, spray second time 7 days intervals, within 7 days, 30 days and 45 days, investigate after second time sprays respectively Gray mold situation.
Investigation method:
Using diagonal five point sampling, each point determines strain 20 strain, samples 100 strain investigation disease indexs altogether.
Fructus Capsici gray mold disease index grade scale:
0 grade: without disease;
1 grade: a small amount of scab occurs in inoculation leaf;
2 grades: lesion area accounts for the 1/3 of leaf area;
3 grades: lesion area accounts for the 1/3-1/2 of leaf area;
4 grades: lesion area accounts for the 1/2-2/3 of leaf area;
5 grades: lesion area accounts for more than the 2/3 of leaf area
Preventive effect computational methods:
Disease index=∑ (morbidity strain number × this morbidity strain level value)/(total investigation strain number × superlative degree value) × 100
Prevention effect (%)=(disease index during initial disease index-investigation)/initial disease index × 100
The prevention effect to Fructus Capsici gray mold used by the different microbial inoculum of table 4
Shown by table 4 result, use this spore microbial inoculum can effectively prevent and treat Fructus Capsici gray mold, after using 7 days, variable concentrations J- 5 spore microbial inoculums to the preventive effect of Fructus Capsici gray mold all more than 80%, suitable with the Routine control prevention effect of chemical pesticide.With often Rule comparison is compared, and J-5 spore microbial inoculum is the most lasting, and after 45 days, preventive effect is all more than 60%.Apparently higher than comparison 28.56%.
By the above results it can be seen that the bacterial strain that preserving number is CGMCC No.11750 that filters out of the present invention is to Fructus Capsici ash Mildew has good antagonism, and Fructus Capsici booth gray mold can effectively be prevented and treated by its spore microbial inoculum.
Bacillus subtilis of the present invention has a suitable prevention effect to Fructus Lycopersici esculenti, Fructus Cucumidis sativi, eggplant, Fructus Capsici and Fructus Fragariae Ananssae, above only For the better embodiment of the present invention, rather than the restriction to its protection domain, those skilled in the art pay creation need not Property work on the premise of any improvement that the present invention is made, be all considered as within the scope of the present invention.
SEQUENCE LISTING
<110>Biology Inst., Hebei Academy of Sciences
<120>a kind of bacillus subtilis J-5 and bacteria agent thereof and application
<130> 2016
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1459
<212> DNA
<213>bacillus subtilis
<400> 1
tcgatatccc tttctttatt acaagctgtc cggcgctttt gccgcatctt gccgtttgtt 60
ttttcagact ggcggtgccc gcgacgtggg ccacctgcct gtcttctggg ataactccgg 120
gaaaccgggg ctaataccgg atggttgttt gaaccgcatg gttcagacat aaaaggtggc 180
ttcggctacc acttacagat ggacccgcgg cgcattagct agttggtgag gtaacggctc 240
accaaggcga cgatgcgtag ccgacctgag agggtgatcg gccacactgg gactgagaca 300
cggcccagac tcctacggga ggcagcagta gggaatcttc cgcaatggac gaaagtctga 360
cggagcaacg ccgcgtgagt gatgaaggtt ttcggatcgt aaagctctgt tgttagggaa 420
gaacaagtgc cgttcaaata gggcggcacc ttgacggtac ctaaccagaa agccacggct 480
aactacgtgc cagcagccgc ggtaatacgt aggtggcaag cgttgtccgg aattattggg 540
cgtaaagggc tcgcaggcgg tttcttaagt ctgatgtgaa agcccccggc tcaaccgggg 600
agggtcattg gaaactgggg aacttgagtg cagaagagga gagtggaatt ccacgtgtag 660
cggtgaaatg cgtagagatg tggaggaaca ccagtggcga aggcgactct ctggtctgta 720
actgacgctg aggagcgaaa gcgtggggag cgaacaggat tagataccct ggtagtccac 780
gccgtaaacg atgagtgcta agtgttaggg ggtttccgcc ccttagtgct gcagctaacg 840
cattaagcac tccgcctggg gagtacggtc gcaagactga aactcaaagg aattgacggg 900
ggcccgcaca agcggtggag catgtggttt aattcgaagc aacgcgaaga accttaccag 960
gtcttgacat cctctgacaa tcctagagat aggacgtccc cttcgggggc agagtgacag 1020
gtggtgcatg gttgtcgtca gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc 1080
gcaacccttg atcttagttg ccagcattca gttgggcact ctaaggtgac tgccggtgac 1140
aaaccggagg aaggtgggga tgacgtcaaa tcatcatgcc ccttatgacc tgggctacac 1200
acgtgctaca atggacagaa caaagggcag cgaaaccgcg aggttaagcc aatcccacaa 1260
atctgttctc agttcggatc gcagtctgca actcgactgc gtgaagctgg aatcgctagt 1320
aatcgcggat cagcatgccg cggtgaatac gttcccgggc cttgtacaca ccgcccgtca 1380
caccacgaga gtttgtaaca cccgaagtcg gtgaggtaac cttttaggag ccagccgccg 1440
aagggggaac aagataagg 1459

Claims (7)

1. a bacillus subtilis J-5, this bacillus subtilis on November 27th, 2015 in Chinese microorganism strain preservation Administration committee's General Microbiological Culture preservation center has carried out preservation, and preserving number is: CGMCC No. 11750.
2. contain the bacteria agent of bacillus subtilis J-5 described in claim 1, it is characterised in that: wherein withered in every gram of microbial inoculum The content of grass bacillus cereus J-5 is 4 × 109cfu/g~7×109cfu。
Bacteria agent the most according to claim 2, it is characterised in that: described microbial inoculum is the spore of bacillus subtilis J-5 The mix bacterium agent of microbial inoculum or spore and thalline.
4. the application in preventing and treating crop gray mold of the bacillus subtilis J-5 described in claim 1.
Application the most according to claim 4, it is characterised in that described crop is Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici or Fructus Fragariae Ananssae.
6. the application in preventing and treating crop gray mold of the bacteria agent described in Claims 2 or 3.
Application the most according to claim 6, it is characterised in that described crop is Fructus Cucumidis sativi, Fructus Lycopersici esculenti, eggplant, Fructus Capsici or Fructus Fragariae Ananssae.
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