CN108004185A - One plant of tool diseases prevention, growth-promoting, drought resisting function plant endogenesis Bei Laisi bacillus and its application - Google Patents

One plant of tool diseases prevention, growth-promoting, drought resisting function plant endogenesis Bei Laisi bacillus and its application Download PDF

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CN108004185A
CN108004185A CN201810017581.8A CN201810017581A CN108004185A CN 108004185 A CN108004185 A CN 108004185A CN 201810017581 A CN201810017581 A CN 201810017581A CN 108004185 A CN108004185 A CN 108004185A
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bacillus
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bei laisi
velezensis
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CN108004185B (en
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郭荣君
王军强
李世东
荆玉玲
高毓晗
孙瑞梁
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Hebei Inuo Biochemical Co., Ltd.
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Institute of Plant Protection of Chinese Academy of Agricultural Sciences
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Abstract

The present invention relates to one plant controlling plant diseases and to have the function of growth-promoting, the plant endogenesis Bei Laisi bacillus of drought resisting and its application.Bei Laisi bacillus (Bacillus velezensis) E6 provided by the present invention is CGMCC No.9665 in the deposit number of China Committee for Culture Collection of Microorganisms's common micro-organisms center.For the strain isolation in plant roots, growth is rapid, plant rhizosphere can rapid, high volume colonize, there is ecological niche advantage；Bacterial strain E6 not only has plant silborne fungal diseases efficient preventive and therapeutic effect, promotes plant root growth, improves plant drought performance, and can destroy the formation of pathogenetic bacteria biofilm, it is not easy to bacterium is produced drug resistance, than other fungicide advantageously；Its nutritional need is simple, production cost is low, has a good application prospect.

Description

One plant tool diseases prevention, growth-promoting, drought resisting function plant endogenesis Bei Laisi bacillus and its Using
Technical field
The invention belongs to agriculture, woods and garden crop protection technique field, and more particularly to one plant prevention plant soil-borne diseases are simultaneously There is the Bei Laisi bacillus of growth-promoting drought resisting and its application.
Technical background
Bacillus is a kind of widely distributed rod-shaped aerobic bacterium of Gram-positive, because its can produce in sprout Spore, has the anti-adversity abilities such as extremely strong heat resistanceheat resistant, radioresistance, anti-chemicals, is easily processed to preparation and is used widely. Commercialization bacillus currently used for control of plant disease is mainly bacillus subtilis formulation.For example, U.S.'s development is withered Careless bacillus GB03, MBI600 and QST713, the bacillus subtilis RB-14 and NB- that Tokyo technical research institute develops 22, bacillus subtilis RC8, bacillus subtilis RC9 and the bacillus subtilis RC11 that Argentina develops, Australia's exploitation Bacillus subtilis A213, bacillus subtilis B903, B908, B916, B1, B3 and BS208 etc. of China's exploitation.In recent years Come, studies have reported that using Bacillus velezensis controlling plant diseases, but it is black to be mainly used for leaf diseases such as Chinese cabbage The prevention of pinta and soil-borne disease such as sclerotinia sclerotiorum.The nearer bacillus amyloliquefaciens with its affiliation, are mainly used for preventing Control graw mold of tomato, rice blast, bacterial leaf streak of rice, tomato wilt and bacterial wilt.And it is directed in production than more serious Cucumber fusarium axysporum, powdery scab of potato, the disease such as bacterial wilt of ginger also lack efficient biocontrol agent, it is normal for bacterial wilt of ginger in production The streptomysin used has made bacterium generate resistance, and copper ion preparation also causes pathogenetic bacteria to generate the cluster of resistance to copper gene, because This obtains the multifunctional efficient Biocontrol Bacillus and its nothing of preventing a variety of soil-borne diseases with China's independent intellectual property right Cell preparation is extremely urgent vital task.
This seminar divided in 2014 out of Plant Protection institute, Chinese Academy of Agricultral Sciences Langfang base marigold root From to one plant of endophytic Bacillus E6, Bei Laisi bacillus velezensis are accredited as, the bacterial strain is to cucumber The multiple diseases such as droop, tomato root rot, powdery scab of potato, bacterial wilt of ginger, soft rot are inhibited, wherein to cucumber Droop and powdery scab of potato have strong inhibitory action, and preventive effect is up to more than 80%, higher than the diseases prevention of other biocontrol microorganisms Effect.The bacterial strain can also improve the drought-resistant ability of plant in addition to prophylaxis effect, and be one plant has application potential very much Bacterial strain.
For plant disease biocontrol microorganisms, the bacteria containing amount in preparation is improved, production cost is reduced, is to realize that biocontrol microorganisms are extensive Production and the precondition of application.The Study on Fermentation in relation to Bacillus velezensis bacterial strains is less both at home and abroad at present, Rarely seen two reports in the country (Wang Zhenhai etc., 2011；Zhen Xinwu and Qiao Junjian, 2013), used in this two researchs Bacillus velezensis bacterial strains are screened from swine excrement, and required suitable fermentation temperature is 37 DEG C, but is come From being 28~32 DEG C in the bacillus E6 suitable growth temperatures in marigold root, other nutritional needs also because its source is different and It is variant.Nutrition is needed also to be not quite similar even if the different strains of same kind.Therefore carry out and have to plant soil-borne diseases There is the Study on Fermentation of efficient prophylaxis effect biocontrol microorganisms Bacillus velezensis E6, development has China's independent intellectual Prevention and control of the biocontrol agent of property right to plant soil-borne diseases have important practical significance.
The content of the invention
The object of the present invention is to provide one plant of prevention plant soil-borne diseases and there is the Bei Laisi buds of growth-promoting and drought resisting Spore bacillus and its application.
Bei Laisi bacillus provided by the present invention is specially Bei Laisi bacillus (Bacillus velezensis) E6 bacterial strains, the bacterial strain are preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms on the 15th in September in 2014 (abbreviation CGMCC, address are the heart：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3), deposit number is CGMCC No.9665.
The bacterium that the present invention also protection activity component is the Bei Laisi bacillus (Bacillus velezensis) E6 Agent.
Absorption carrier is may also include in the microbial inoculum；The absorption carrier can be diatomite, turf, vermiculite, powdered rice hulls, jade At least one of rice core powder, perlite and calcium carbonate.
More specifically, in one embodiment of the invention, the absorption carrier is specifically pressed by powdered rice hulls and diatomite Formed according to the proportioning mixing of volume ratio 2: 1.
Further, the microbial inoculum by the Bei Laisi bacillus (Bacillus velezensis) E6 zymotic fluid, or Gained bacterium mud is mixed with the absorption carrier according to the proportioning of 1L: 1Kg to 1L: 3Kg (such as 1L: 1Kg) after being centrifuged to zymotic fluid With into.
In one embodiment of the present of invention (embodiment 2), the Bei Laisi bacillus (Bacillus is prepared Velezensis bulk fermentation nutrient solution) is used during the zymotic fluid of E6, it is described in the zymotic fluid of the Bei Laisi bacillus E6 The viable count of Bei Laisi bacillus E6 is 80~13,000,000,000/mL (such as 8,000,000,000/mL) described zymotic fluid.Wherein, a large amount of hairs The solvent of ferment nutrient solution is water, and solute and concentration are as follows：Sucrose 0.4~0.8%, cornstarch 1.5~1.8%, beancake powder 1.2~1.5% and calcium carbonate 0.2~0.5%；Wherein, % represents mass percentage.Correspondingly, in preparation-obtained institute State in microbial inoculum, the viable count of the Bei Laisi bacillus E6 is 40~100,000,000,000/g (such as 100,000,000,000/g) described microbial inoculum；It is described The water content of microbial inoculum is within 5~8%.
The preparation method of the microbial inoculum falls within protection scope of the present invention.
The preparation method of the microbial inoculum, specifically may include following steps：
(1) Bei Laisi bacillus (Bacillus velezensis) E6 is cultivated, obtains the Bei Laisi gemma bar The zymotic fluid of bacterium E6；In the zymotic fluid, the viable count of the Bei Laisi bacillus for 80~13,000,000,000/mL (such as 8,000,000,000/ mL)；
(2) by zymotic fluid obtained by step (1) or the bacterium mud after centrifugation and the absorption carrier according to 1L: 1Kg to 1L: 4Kg The proportioning of (such as 1L: 1Kg) mixes, and obtains the microbial inoculum.
In step (1), cultivate the Bei Laisi bacillus (Bacillus velezensis) E6 culture medium it is molten Agent is water, and solute and concentration are as follows：Sucrose 0.4~0.8%, cornstarch 1.5~1.8%, beancake powder 1.2~1.5% and carbon Sour calcium 0.2~0.5%；Wherein, % represents mass percentage.
More specifically, the solvent of the culture medium is water, and solute and concentration are as follows：Sucrose 0.5%, cornstarch 1.7%th, beancake powder 1.4% and calcium carbonate 0.45%；Wherein, % represents mass percentage.
The zymotic fluid of Bei Laisi bacillus (Bacillus velezensis) E6 falls within the protection model of the present invention Enclose.
It is used to prevent plant soil-borne diseases, promote plant growth or improve plant drought performance present invention also offers one kind Product.
The product provided by the present invention for being used to prevent plant soil-borne diseases.
Bei Laisi bacillus (Bacillus velezensis) the E6 or described microbial inoculums or the zymotic fluid are as follows (1) or the application in (2) falls within protection scope of the present invention：
(1) plant soil-borne diseases are prevented, promote plant growth or improve plant drought performance；
(2) product for being used for preventing plant soil-borne diseases is prepared
The plant soil-borne diseases are specially by Fusarium oxysporum (Fusarium oxysporum), Fusarium solani (Fusarium solani), rhizoctonia (Rhizoctonia solani), powdery scab bacterium (Spongospora Subterranea), ralstonia solanacearum (Ralstonia solanacearum), Bacteria erwinia (Pectobacterium Chrysanthemi, Erwinia carafavora) etc. disease fungus and bacterial soil-borne disease.
The plant concretely cucumber, tomato, potato, ginger, konjaku, wheat.
Present invention also offers a kind of method for preventing plant soil-borne diseases.
The method of prevention plant soil-borne diseases provided by the present invention is concretely following (A), (B), (C) or (D)：
(A) include the following steps：The microbial inoculum dilution is added into vegetable seeds, is sowed after being dressed seed.Described The concentration of microbial inoculum dilution is 0.1 hundred million/mL；
(B) include the following steps：The root system of the plant is placed in the dilution of the microbial inoculum and is soaked, is planted after immersion Kind, and in growth period pouring root.The viable count of the microbial inoculum dilution is 0.1 hundred million/mL；
(C) include the following steps：The microbial inoculum is mixed with seedling medium, then by the plant seeding in seedling medium In；
(D) above-mentioned (A), (B), any combination two-by-two of (C) method or being applied in combination for three kinds of methods.
In (A), the viable bacteria of Bei Laisi bacillus described in the microbial inoculum (Bacillus velezensis) E6 Number is 4,000,000,000/g；The vegetable seeds and the microbial inoculum be (Bei Laisi bacillus (Bacillus velezensis) E6's Viable count is 4,000,000,000/g) mixing proportioning concretely 1: 1 (mass ratio).Further, described dress seed is specially：By the microbial inoculum (viable count of Bei Laisi bacillus (Bacillus velezensis) E6 is 4,000,000,000/g) is with water according to 1g: 400mL Proportioning be diluted to scattered paste shape thing, then add be stirred with the vegetable seeds of the quality such as the microbial inoculum thereto.
In (B), Bei Laisi bacillus described in the dilution of the microbial inoculum (Bacillus velezensis) The viable count of E6 is 0.1 hundred million/mL；The time of the immersion is 30min.
In (C), Bei Laisi bacillus described in matrix (Bacillus velezensis) E6 after the mixing Viable count be 0.1 hundred million/g；
In (D), the concentration of the microbial inoculum is consistent with respective concentration in (A), (B), (C)..
Bei Laisi bacillus (Bacillus velezensis) E6CGMCC No.9665 provided by the present invention have Following advantage：
1st, the strain isolation is in plant roots, being suitable for plant rhizosphere environment, and grow it is rapid, can be in plant roots Border rapid, high volume colonizes, and has ecological niche advantage.
2nd, currently used bacterium fungicide or copper ion fungicide are easily led to bacterial resistance occurred after long-term use, And increase the difficulty of disease control.And the sterile ferment filtrate of the bacterial strain can significantly inhibit pathogenetic bacteria ralstonia solanacearum and Bacteria erwinia The formation of biomembrane, is by disturbing signal transmission between bacterium and envelope that outside is wrapped up inhibits bacteria, is a kind of The new mode of action, this mode of action are not easy to make pathogenetic bacteria develop immunity to drugs, and more have than other bactericidal effect modes excellent Gesture, the bacterial strain are one plant of potential Antagonistic Fungis.
3rd, there is efficient preventive and therapeutic effect to plurality of plant diseases, moreover it is possible to promote plant growth and improve the drought resistance of plant Energy.
4th, the nutritional need of the bacterial strain is fairly simple, and fermentation period is short, and cost is low, and easily large-scale production and commercialization should With.
Bei Laisi bacillus (Bacillus velezensis) E6CGMCC No.9665 provided by the present invention are One plant of endogenetic bacteria is separated to out of Plant Protection institute, Chinese Academy of Agricultral Sciences Langfang base marigold root within 2014.Should The characteristics of bacterial strain is：When being cultivated on nutrient medium tablet, bacterium colony projection forms gauffer, and quickly, this is bacterium colony expansion rate One of precondition that biocontrol microorganisms can be mass-produced and apply.To cucumber fusarium axysporum, tomato root rot, powdery scab of potato etc. Multiple diseases are inhibited.The bacterial strain can also improve the drought resisting function of plant in addition to prophylaxis effect, be one plant Very potential bacterial strain.Identified, which is Bei Laisi bacillus velezensis, is named as E6.Should Bacterial strain has cucumber fusarium axysporum and powdery scab of potato strong inhibitory action, and preventive effect is up to more than 80%, higher than other lifes Fungi-proofing protection effect.And growth can be inhibited bacteria by destroying the formation of pathogenetic bacteria biofilm, avoid largely using Pathogenetic bacteria may be caused to produce the probability of drug resistance.During E6 strain fermentation cultures, when the speed of growth is fast, 28 is small within can complete Fermentation, viable count can reach 80~13,000,000,000/mL after fermentation, low into production cost.Therefore E6 is one plant of very potential plant Thing biocontrol bacteria in soil-borne diseases, so as to propose the present invention.
Preservation explanation
Strain name：Bei Laisi bacillus
Latin name：Bacillus velezensis
Strain number：E6
Preservation mechanism：China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is referred to as：CGMCC
Address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date：On September 15th, 2014
Collection is registered on the books numbering：CGMCC No.9665
Brief description of the drawings
Fig. 1 is subject to upgrowth situation figure of the drought stress after 1 day for wheat
Embodiment
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples, is commercially available unless otherwise specified.
The separation and identification of embodiment 1, Bei Laisi bacillus (Bacillus velezensis) E6
(1) separation of bacillus E6 bacterial strains
Bei Laisi bacillus (Bacillus velezensis) E6 of the present invention is out of marigold root, and tissue is ground Grinding fluid separates what is obtained after being diluted coating, and separation method is：
(1) separation of Bei Laisi bacillus：In Langfang City academy of agricultural sciences of Hebei province proving ground, the plant such as collection marigold Root.1g roots are weighed, after 1%NaClO disinfection washings, is put into sterile mortar and grinds, 1mL is drawn from mortar with liquid-transfering gun Lapping liquid, adds in another test tube for filling 9mL sterile waters, is mixed evenly, and so on be respectively prepared 10-3、10-4、10-5No With the solution of dilution factor.100 μ L dilutions are taken to be coated on TSA culture medium flat plates, three culture dishes of each gradient coating, 27~ 30 DEG C are cultivated 2~4 days.The bacterium of picking different shape, is transferred on nutrient agar (i.e. NA solid mediums) and rules Purifying.
Wherein, the TSA nutrient media componentses and proportioning are：Tryptone 15g, soy peptone 5g, sodium chloride 5g, agar 15g, distilled water 1000mL, adjust pH to 7.0,121 DEG C of autoclavings 30 minutes.
The NA solid mediums component and proportioning are：Glucose 10g, peptone 10g, beef extract 3g, yeast extract 1g, fine jade Fat 18g, distilled water 1000mL, adjust pH to 7.0,121 DEG C of autoclavings 30 minutes.
(2) inhibitory action of the bacillus E6 to disease fungus
By disease funguses such as the Fusarium oxysporum activated, Fusarium solani, Rhizoctonia solani Kuhns, it is made of 5mm card punch Pure culture biscuits involvng inoculation is central in PDA tablets, and the Bei Laisi bacillus E6,3 repetitions, to be only inoculated with are connect away from point at bacteria cake 2cm Pathogen is used as blank control.3~4 days measurement colony diameters are cultivated at 26 DEG C, calculate colony growth inhibiting rate.As a result table Bright, bacillus E6 has inhibitory action to above-mentioned disease fungus.
Inhibitory action of the 1 bacillus E6 of table to disease fungus
(3) inhibitory action of the bacillus E6 to pathogenetic bacteria
Influence to pathogenetic bacteria growth：Take out, 4 DEG C of refrigerators save backup.
Ralstonia solanacearum, Bacteria erwinia and Bei Laisi bacillus E6 bacterial strains are transferred to the training of 5mL beef extract-peptones liquid In nutrient solution, 30 DEG C, 180 revs/min of shake cultures, ralstonia solanacearum and Bacteria erwinia cultivate 12~18 it is small when after, blood cell counting plate Count, adjustment pathogen bacteria suspension concentration is 108/mL.100 μ L Bacteria culturing liquid are drawn, are added to beef extract-peptone training Support on base tablet, coating, dries.Bei Laisi bacillus E6 is transferred in centrifuge tube after 48h is cultivated, 4 DEG C, 10000 turns/ Point, centrifuge 10 minutes, supernatant crosses 0.22 μm of filter membrane, spare.Oxford cup is placed on the culture medium of coating pathogen, added The sterile ferment filtrate of 200 μ L Bei Laisi bacillus E6, each pathogen do the repetition of 3 tablets.Culture dish is placed on 30 DEG C Cultivated in incubator, after 24~48h, vernier caliper measurement antibacterial circle diameter (mm).The result shows that add bacillus E6 fermentations Obvious inhibition zone is produced around the Oxford cup of filtrate, diameter is more than 16mm.
The inhibitory action formed to pathogenetic bacteria biofilm：
150 μ L pathogenetic bacterias suspension are made an addition in flat 96 hole polystyrene Tissue Culture Plate respectively, are then added The bacteria-free filtrate of 50 μ L Bei Laisi bacillus E6, when being placed in that quiescent culture 2 is small in 28~30 DEG C of incubators, will be trained with liquid-transfering gun Nutrient solution and it is not adhered to the pathogenetic bacteria of culture plate and sops up, is gently rinsed twice, be finally adhered to thin with phosphate buffer What born of the same parents cultivated plate surface is the initial bacterium colony of biofilm, then adds the beef extract-peptone liquid medium of 200 μ L, is stood Culture is formed to biofilm for 24~36 hours.Observe thalline turbidity and surface filming situation.The result shows that bacterial strain E6 is sterile Ferment filtrate significantly inhibits the formation of pathogenetic bacteria biofilm, and inhibiting rate is respectively 43.0% and 45.8%.
(4) identification of bacillus E6 bacterial strains
The genomic DNA of bacterial strain E6 is extracted, expands 16S rDNA sequences with universal primer 27f and 1492r, and surveyed Sequence.
27f：5′-AGA GTT TGA TCC TGG CTC AG-3′；1492r：5′-TAC GGC TAC CTT GTTACG ACT T-3′。
The sequence of the 16S rDNA of bacterial strain E6 submitted GenBank, accession number MG550967.In view of above-mentioned molecule life Thing CHARACTERISTICS IDENTIFICATION by the bacterial strain E6 that step 1 separates and purifies as a result, be accredited as Bei Laisi bacillus (Bacillus velezensis).The Bei Laisi bacillus (Bacillus velezensis) is preserved in China on 15th in September in 2014 (abbreviation CGMCC, address are Microbiological Culture Collection administration committee common micro-organisms center：BeiChen West Road, Chaoyang District, BeiJing City 1 Number institute 3), deposit number is CGMCC No.9665.
Embodiment 2：
The preparation of Bei Laisi bacillus (Bacillus velezensis) E6 microbial inoculums
1st, Bei Laisi bacillus (Bacillus velezensis) the E6CGMCC No.9665's obtained embodiment 1 Lawn is transplanted in nutrient broth (NB), and 28~30 DEG C of shaking table shaken cultivation 20h obtain seed liquor.
Wherein, the formula of nutrient broth (NB) is：Glucose 10g, peptone 10g, beef extract 3g, yeast extract 1g, distillation Water 1000mL, pH are adjusted to 7.0~7.2,121 DEG C of autoclavings 30 minutes.
2nd, the seed liquor for obtaining step 1 in 1% ratio (volume ratio) is inoculated into mass propgation fermentation culture, culture 30~32 DEG C of temperature, 150~180 revs/min of rotating speed, initial pH 7.0~7.2, incubation time 28h.Fermented after the completion of fermentation Liquid, the viable count of Bei Laisi bacillus (Bacillus velezensis) E6CGMCC No.9665 is up to 120 in zymotic fluid Hundred million/mL.
3rd, the bacterium mud and absorption carrier --- rice husk obtained the Bei Laisi fermentation of bacillus liquid that step 2 obtains through centrifugation Powder and diatomite (volume ratio 2: 1) are mixed evenly according to the ratio that volume mass ratio is 1: 1 (i.e. 1L: 1Kg), up to microbial inoculum.Institute Centrifugal rotational speed is stated as 11500 revs/min, centrifugation time is 30 minutes；Bei Laisi bacillus (Bacillus in the microbial inoculum Velezensis) viable count of E6CGMCC No.9665 is up to 100,000,000,000/g, and water content is within 5%.
Embodiment 3：The prophylaxis effect of bacillus E6 microbial inoculums
To the prevention effect of cucumber fusarium axysporum：First sickle-like bacteria is cultivated with Armstrong nutrient solutions, mirror after 3~4 days Inspection counts, and bacterium solution is diluted to 5 × 104Cfu/mL is spare；E6 microbial inoculums are diluted to 1 × 107cfu/mL.By the spore of sickle-like bacteria Suspension mixes in equal volume with E6 microbial inoculums, then adds in mixed liquor the seed disinfected and soaks seed 30 minutes.Often handle 10 Repeat, each 3 cucumber seeds of repeat sowing, are compared with not applying the processing of microbial inoculum.The growth of cucumber is investigated after sowing 3 weeks Incidence.
The component and proportioning of the Armstrong nutrient solutions be：Glucose 20.0g, KCl 1.6g, MgSO4.7H2O 0.4g、Ca(NO3)2 5.9g、KH2PO41.1g, FeSO40.2 μ g/mL, ZnSO40.2μg/mL、MnSO40.2 μ g/mL, distilled water 1000mL, 121 DEG C of autoclaving 30min.
Wherein, FeSO4、ZnSO4、MnSO4Mother liquor can be first made into.Above-mentioned three kinds of medicine 0.02g are weighed respectively in 100mL In distilled water, 1mL is taken after mixing in above-mentioned 999mL mixed liquors.
Investigation result (table 2) shows that E6 microbial inoculums are to the preventive effect of cucumber fusarium axysporum up to 84.2%.
Protection effect of the 2 E6 microbial inoculums of table to cucumber fusarium axysporum
To the prevention effect of tomato root rot：
The sick soil of rhizoctonia is collected, mixes, is then charged into according to the ratio of mass ratio 1: 1 with sterile seedling medium In the square seedling-raising box of 16.5cm × 11.5cm.About 4cm one wide, the ditch of deep 6cm are excavated in sick soil.By E6 microbial inoculums and nothing Bacterium matrix mix, make its final concentration of 106Microbial inoculum, is then sprinkled into ditch by cfu/g, and tomato seedling is transplanted in ditch, and with dilute Microbial inoculum envelope ditch after releasing, about needs to spread fertilizer over the fields microbial inoculum 20g per bar ditch.The repetition of 3 flowerpots is often handled, per 10 young plant of pot transplanting, is moved Hot-house culture is put into after cultivation, temperature is 32 DEG C of daytime, 22 DEG C at night.The Investigate incidence after transplanting 9 days, calculates protection effect. As a result (table 3) shows, E6 microbial inoculums are to the preventive effect of tomato root rot up to 69.3%.
3 sick soil method of table measures protection effect of the E6 microbial inoculums to tomato root rot
To the prevention effect of powdery scab of potato and early blight：
Seed potato is handled with E6 microbial inoculums, dosage is 1000g/ mus, compares and dresses seed for thiophanate methyl, and dosage is 300g/ mus, potato wedge investigation is dug out at the end of growth period, it is found that the potato wedge surface after the processing of E6 microbial inoculums is smooth, powdery scab of potato Disease index substantially mitigate, preventive effect is up to 86.58%；Per mu yield increases by 17.2% (table 4).
4 E6 microbial inoculums potato seed of table handles the preventive and therapeutic effect to powdery scab of potato
Embodiment 4：The growth-promoting drought resisting function of bacillus E6
Take field native, be attached in flowerpot.Wheat seed is seeded in soil after being soaked seed with microbial inoculum.E6 bacteria suspension concentrations are 108Cfu/g, 20 wheats are sowed per basin, often handle 5 repetitions.Wheat seed not soak seed is used as control CK.In sowing Afterwards 3 weeks when carry out water shortage and handle 2 days, then rewater, Plant Height in Wheat, root fresh weight and strain fresh weight measured after 3 days.
The survey showed that：E6 microbial inoculums handle wheat seed, can promote plant strain growth (table 5), well developed root system；It is subject to arid After stress, well-grown, color is dark green, and compares yellow leaf (Fig. 1).
Influence of the 5 E6 microbial inoculums of table to wheat seeds sprouting and growth

Claims (9)

  1. Bei Laisi bacillus 1. (Bacillus velezensis) E6, is stored in Chinese microorganism strain preservation conservator Meeting common micro-organisms center, deposit number is CGMCC No.9665.
  2. 2. a kind of microbial inoculum, its active ingredient is the Bei Laisi bacillus (Bacillus described in claim 1 velezensis)E6。
  3. 3. microbial inoculum according to claim 2, it is characterised in that：In the microbial inoculum, the Bei Laisi bacillus The viable count of (Bacillus velezensis) E6 is the liquid bacterial agent of 80~13,000,000,000/mL.
  4. 4. the microbial inoculum according to Claims 2 or 3, it is characterised in that：The microbial inoculum is by the Bei Laisi bacillus The zymotic fluid of (Bacillus velezensis) E6 is directly or the bacterium mud after centrifugation and absorption carrier are according to 1L: 1Kg to 1L: The proportioning of 3Kg mixes the solid fungicide formed.
  5. 5. the preparation method of microbial inoculum described in claim 4, includes the following steps：
    (1) Bei Laisi bacillus (Bacillus velezensis) E6 described in claim 1 is cultivated, obtains the shellfish Lay The zymotic fluid of this bacillus (Bacillus velezensis) E6；
    (2) absorption carrier described in bacterium mud and claim 2 after centrifugation of zymotic fluid or zymotic fluid obtained by step (1) is pressed Mixed according to 1L: 1Kg to 1L: 3Kg proportioning, obtain the microbial inoculum.
  6. 6. the culture medium prescription in the step of bacterial preparation process according to claim 5 (1), the Bei Laisi gemma bar The solvent of the nutrient solution of bacterium (Bacillus velezensis) E6 is water, and solute and concentration are as follows：Sucrose 0.4~0.8%, jade Rice starch 1.5~1.8%, beancake powder 1.2~1.5% and calcium carbonate 0.2~0.5%；Wherein, % represents mass percentage.
  7. 7. prepared by Bei Laisi bacillus (Bacillus velezensis) E6 described in claim 1 is used to prevent plant Soil-borne disease, promote plant growth and improve the product of plant drought performance.
  8. 8. tool diseases prevention, growth-promoting, drought resisting are multi-functional as described in claim 2, claim 3, claim 4 and claim 7 Bei Laisi bacillus (Bacillus velezensis) E6 microbial inoculums, zymotic fluid, the purposes of solid fungicide and product, particularly The prevention of the plant soil-borne diseases caused by pathogen infection.The plant soil-borne diseases are by sickle-like bacteria Fusarium Oxysporum, Fusarium solani, rhizoctonia Rhizoctonia solani, powdery scab bacterium Spongospora Subterranea, ralstonia solanacearum Ralstonia solanacearum, Bacteria erwinia Pectobacterium The disease funguses such as chrysanthemi, Erwinia carafavora and bacterial soil-borne disease.
  9. 9. Bei Laisi bacillus (Bacillus velezensis) E6 fermentation generations as described in claim 1 and claim 2 Thank to thing and produce the application for the treatment of of relevant gene and antibacterial material in agricultural, animal husbandry and biomedicine field.
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