CN104073451A - Paenibacillus polymyxa for resisting multiple peony pathogenic bacteria and application thereof - Google Patents
Paenibacillus polymyxa for resisting multiple peony pathogenic bacteria and application thereof Download PDFInfo
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Abstract
The invention discloses paenibacillus polymyxa for resisting multiple peony pathogenic bacteria and an application of paenibacillus polymyxa. The paenibacillus polymyxa for resisting multiple peony pathogenic bacteria is paenibacillus polymyxa CSM1101, and the preservation number of paenibacillus polymyxa in the China General Microbiological Culture Collection Center is CGMCC No.8527. The paenibacillus polymyxa CSM1101 can remarkably inhibit Botrytis paeoniae, Cladosporium paeoniae and Cercospora varricolor Winter, and can remarkably promote growth of peony.
Description
Technical field
The present invention relates to Paenibacillus polymyxa and the application thereof of the multiple tree peony pathogenic bacteria of a strain antagonism.
Background technology
Tree peony returns Paeoniaceae (Paeoniaceae), Paeonia (Paeonia), is all one of Chinese most important ornamental plant and medicinal plant all the time, the history of existing more than 1600 year.At present, tree peony Main Cultivation place is at home Heze, Luoyang, Beijing, Linxia, day Peng, Tongling etc.In recent years, the oil of tree peony day by day highlights with being worth, and oil presents the situation of Rapid Expansion with the development of tree peony.For viewing and admiring tree peony, distinct issues are, due to the soil micro-ecosystem activity decreased that the difference of different areas climatope and soil property causes, are to introduce a fine variety one of major reason that regional culture tree peony upgrowth situation is bad, fancy points declines.And for oil with tree peony, how to apply fertilizers scientifically and the output that significantly improves peony seeds is very urgent problem.In the cultivation process of tree peony, be also faced with the threat of some fungal diseases, mainly contain gray mold (Botrytis paeoniae), red spot disease (Cladosporium aeoniae) and brown spot (Cercospora paeoniae) etc.Therefore, development promotes the microbiobacterial agent of tree peony growth antagonism pathogenic bacteria to have remarkable and urgent realistic meaning.Wherein, the plant-growth promoting rhizobacteria microorganism resource that screening has growth-promoting and antagonism pathogenic bacteria function concurrently to tree peony is the important bottleneck factor of restriction micro-organisms microbial inoculum development.
Summary of the invention
A technical problem to be solved by this invention is to provide the multiple tree peony pathogenic bacteria of a strain energy antagonism, promotes tree peony growth and can take the bacterial strain of maize straw as raw material production promotion tree peony growth antagonism pathogenic bacteria.
Bacterial strain provided by the present invention is Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101, on December 09th, 2013, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center (being called for short CGMCC), preservation registration number is CGMCC No.8527.
The morphological feature of described Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 is as follows: Gram-positive, and motion, it is shaft-like that thalline is, and diameter is 0.6-0.8 μ m, and long is 2.0-7.6 μ m.Gemma column, 1.6-3.5 * 1.2-1.5 μ m, middle life is raw to time end.Sporocyst obviously enlarges into spindle-type or bar-shaped.The irregular splintery of its colony edge, translucent to opaque.Rear its bacterium colony is coarse, shallow white, middle slightly projection.Bacterium colony adheres to media surface.
The physiological and biochemical property of described Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 is as follows:
Hydrolyzed starch: feminine gender;
Caseinhydrolysate: feminine gender;
Gelatin hydrolysate: feminine gender;
Utilize Citrate trianion: feminine gender;
Tyrosine decomposes: feminine gender;
Phenylalanine deaminase experiment: feminine gender;
Yolk lecithin enzyme experiment: feminine gender;
Indoles produces: feminine gender;
Hippurate experiment: feminine gender;
Catalase experiment: the positive;
Nitrate reduction experiment: the positive;
Protosol produces: the positive;
Anti-N,O-Diacetylmuramidase experiment: the positive;
Decomposition glucose produces sour aerogenesis: the positive;
Decompose pectinose and produce sour aerogenesis: the positive;
Decompose wood sugar and produce sour aerogenesis: the positive;
Decompose N.F,USP MANNITOL and produce sour aerogenesis: the positive;
Salt tolerance test: 7%NaCl growth;
In the nutrient broth medium (NB) of pH6.8 and the Sharpe glucose broth of pH5.7, all can grow.On MR-VP meat soup, produce acetyl methyl carbinol, pH6.0.
The 16SrDNA sequence of described Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 is as shown in SEQ ID No.1.Wherein, SEQ ID No.1 is comprised of 1496 Nucleotide.
Another object of the present invention is to provide a kind of microbial inoculum, and the activeconstituents of this microbial inoculum is described Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101.
This microbial inoculum, except Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 comprising as activeconstituents, also can comprise auxiliary material, as the stalk of the ight soil of the peat composed of rotten mosses, animal, all kinds of crops, loose shell, straw, peanut skin etc.Described microbial inoculum also can comprise carrier.Described carrier can be solid carrier or liquid vehicle.Described solid carrier is mineral material, vegetable material or macromolecular compound; Described mineral material is at least one in clay, talcum, kaolin, montmorillonite, white carbon, zeolite, silica and diatomite; Described vegetable material is at least one in Semen Maydis powder, bean powder and starch; Described macromolecular compound is polyvinyl alcohol and/or polyglycol.Described liquid vehicle can be water.In described microbial inoculum, described activeconstituents can exist with viable cell, the fermented liquid of viable cell, the form of the mixture of the filtrate of cell culture or cell and filtrate of being cultivated.The formulation of described composition can be multiple formulation, as liquor, emulsion, suspension agent, pulvis, granule, wettable powder or water dispersible granules.
Described Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 can be following A 1)-A3) in any microbial inoculum:
A1) promote the microbial inoculum of tree peony growth and antagonism pathogenic bacteria;
Described pathogenic bacteria be in tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) three kinds, any two or any.
A2) microbial inoculum of antagonism pathogenic bacteria; Described pathogenic bacteria be in tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) three kinds, any two or any.
A3) promote the microbial inoculum of tree peony growth.
Described Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 preparation described A1)-A3) and in any microbial inoculum in application also belong to protection scope of the present invention.
The following purposes of above-mentioned Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 or above-mentioned microbial inoculum also belongs to protection scope of the present invention:
C1) promote tree peony growth and antagonism pathogenic bacteria;
C2) described promotion tree peony growth;
C3) described antagonism pathogenic bacteria.
The present invention also provides a kind of method that promotes tree peony growth.
The method of promotion tree peony provided by the present invention growth, is included in before taking root the Paeonia suffruticosa seed after seed soaking is processed to the step that 1-3 hour (as 1 hour) carries out root culture again with described microbial inoculum.
In aforesaid method, by the Paeonia suffruticosa seed after seed soaking with the mode of described microbial inoculum processing because of the physical condition of described microbial inoculum different, if described microbial inoculum is liquid, directly with described microbial inoculum, soak the Paeonia suffruticosa seed after described seed soaking, the content of the described Paenibacillus polymyxa in described microbial inoculum (Paenibacillus polymyxa) CSM1101 can be 10
8cfu/ml-10
9cfu/ml(is as 10
9cfu/ml); If described microbial inoculum is solid-state, need to make liquid bacterial agent to adding water in described microbial inoculum, with this liquid bacterial agent, soak the Paeonia suffruticosa seed after described seed soaking again, the content of the described Paenibacillus polymyxa in described liquid bacterial agent (Paenibacillus polymyxa) CSM1101 can be 10
8cfu/ml-10
9cfu/ml(is as 10
9cfu/ml).
In a specific embodiment of the present invention, described tree peony is Feng Dan (Paeonia ostii).
In above-mentioned literary composition, the growth of described promotion tree peony can be presented as following B1)-B7) in any:
B1) shorten Paeonia suffruticosa seed rootage duration;
B2) improve Paeonia suffruticosa seed rooting rate;
B3) shorten the germinating time of Paeonia suffruticosa seed;
B4) improve the percentage of germination of Paeonia suffruticosa seed;
B5) improve the main root length of tree peony;
B6) improve tree peony height of seedling;
B7) improve tree peony Seedling Biomass;
Described raising tree peony Seedling Biomass is presented as fresh weight or the dry weight that improves tree peony seedling.
In aforesaid method, described pathogenic bacteria can be in tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) three kinds, any two or any.
Another object of the present invention is to provide the method for a kind of cultivation Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101.
The method of cultivation Paenibacillus polymyxa provided by the present invention (Paenibacillus polymyxa) CSM1101, comprises Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 in the step of cultivating for cultivating the substratum of Paenibacillus polymyxa.
Another object of the present invention is to provide a kind of method of preparing described microbial inoculum.
The method of the described microbial inoculum of preparation provided by the present invention, comprises the steps: described Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 as activeconstituents, to obtain described microbial inoculum.
Paenibacillus polymyxa of the present invention (Paenibacillus polymyxa) CSM1101 can significantly suppress tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter), and their bacteriostasis rate has been reached respectively to 57.2%, 65.7% and 88.3%.The rootage duration of the Paeonia suffruticosa seed (processing of microbial inoculum group lamination) that Paenibacillus polymyxa of the present invention (Paenibacillus polymyxa) CSM1101 processes is processed and has been shifted to an earlier date 10 days than control group lamination; The rooting rate that microbial inoculum group lamination is processed reaches 95.0%, and the rooting rate of processing (not processing with microbial inoculum) than control group lamination has improved 33.8%; The main root length that microbial inoculum group lamination is processed is 69.0mm, is 1.68 times that control group lamination is processed; The seed percentage of main root length >=40mm that microbial inoculum group lamination is processed reaches 90 ± 0.5%, is 1.3 times that control group lamination is processed; The germinating time that microbial inoculum group lamination is processed is processed and has been shortened 10 days than control group lamination; The percentage of germination that microbial inoculum group lamination is processed reaches 99.0%, is 1.2 times that control group lamination is processed; The height of seedling that microbial inoculum group lamination is processed is 1.77 times that control group lamination is processed, and the dry weight that microbial inoculum group lamination is processed is 2.27 times that control group lamination is processed.Explanation take promotion tree peony growth that Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 is activeconstituents antagonism pathogenic bacteria microbial inoculum not only antagonism tree peony pathogenic bacteria also promoted significantly taking root (radicle sprouting) of Paeonia suffruticosa seed and germinateed (plumule sprouting) and the growth of tree peony seedling.
preservation explanation
Strain name: Paenibacillus polymyxa
Latin name: Paenibacillus polymyxa
Strain number: CSM1101
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is called for short: CGMCC
Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City
Preservation date: on December 09th, 2013
The preservation center numbering of registering on the books: CGMCC No.8527
Accompanying drawing explanation
Fig. 1 is that microbial inoculum group lamination is processed and the control group lamination treating part seed root culture photo of 60 days.
A is that control group lamination is processed, and b is that microbial inoculum group lamination is processed.
Embodiment
Below in conjunction with embodiment, the present invention is further described in detail, the embodiment providing is only in order to illustrate the present invention, rather than in order to limit the scope of the invention.Experimental technique in following embodiment, if no special instructions, is ordinary method.In following embodiment, material used, reagent etc., if no special instructions, all can obtain from commercial channels.
Tree peony botrytis (Botrytis paeoniae) ACCC36053 and tree peony branch spore mould (Cladosporium paeoniae) ACCC36063 used in following embodiment were all concealed in China Committee for Culture Collection of Microorganisms agricultural microorganism center (abbreviation ACCC before the application's the applying date, address: No.12 ,zhongguancun south street,Haidian District, Beijing, INST OF AGRICULTURAL RESOURCES, postcode 100081), be on August 31st, 2006 the collection day of these two bacterial strains, from collection, the public can obtain this bacterial strain from Chinese microorganism strain preservation management committee agricultural microorganism center.
In following embodiment, tree peony Pathogenic Bacteria Causing Brown Blotch Disease used is the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) (Zhang Jianguo, the Major Diseases of 2001. tree peonies and control. Chinese flower gardening, 23:32-34) public can obtain from Shanghai Chenshan Botanical Garden, this biomaterial related experiment of the present invention of only attaching most importance to is again used, not can be used as other purposes and uses.
Tree peony in following embodiment is Feng Dan (Paeonia ostii) (Jigang Han, Yao Song, Zhigang Liu, etc.2011.Culturable bacterial community analysis in the root domainsof two varieties of tree peony (Paeonia ostii) .FEMS Microbiol Lett, 322:15 – 24) public can obtain from Shanghai Chenshan Botanical Garden, this biomaterial related experiment of the present invention of only attaching most importance to is again used, not can be used as other purposes and uses.
Substratum used in following embodiment is as follows:
Semi-solid
nitrogen-free agar: sucrose, 10g; Oxysuccinic acid, 5g; KH
2pO
4h
2o, 0.4g; K
2hPO
4h
2o, 0.1g; MgSO
47H
2o, 0.2g; NaCl, 0.1g; CaCl
22H
2o, 0.02g; FeCl
3,0.01g; Na
2moO
42H
2o, 0.02g; Agar, 3g; With distilled water, be settled to 1000ml, pH7.0-7.2.121 ℃ of vapor sterilization 20min.
nitrogen-free agar: sucrose, 10g; Oxysuccinic acid, 5g; KH
2pO
4h
2o, 0.4g; K
2hPO
4h
2o, 0.1g; MgSO
47H
2o, 0.2g; NaCl, 0.1g; CaCl
22H
2o, 0.02g; FeCl
3,0.01g; Na
2moO
42H
2o, 0.02g; Agar 18g; With distilled water, be settled to 1000ml, pH7.0-7.2.121 ℃ of vapor sterilization 20min.
NB substratum: glucose 5.0g, peptone 5.0g, extractum carnis 3.0g, is settled to 1000ml with distilled water, pH7.0-7.2,121 ℃ of sterilizing 20min.
NA substratum: glucose 5.0g, peptone 5.0g, extractum carnis 3.0g, agar 18g, is settled to 1000ml with distilled water, pH7.0-7.2,121 ℃ of sterilizing 20min.
PDA substratum: glucose 20g, agar 18g, potato 200g, adds water 1000mL and boils 30min leaching juice, adds water and supplies 1000ml, pH6.0-7.0,121 ℃ of sterilizing 20min.
Cellulase in following embodiment is purchased from raw work biotechnology (Shanghai) limited-liability company.Cellulose enzyme unit of activity (FPU) in following embodiment is defined as 1g cellulase powder and at 55 ℃, in 60min, is hydrolyzed filter paper gained glucose μ mol number.The concrete measuring method of cellulose enzyme vigor is as follows: extracting cellulose enzyme 10.00g, 0.05mol/L acetic acid-the sodium-acetate buffer that adds 100mlpH4.85 fully dissolves, obtain cellulase solution, by after cellulase solution dilution, draw 0.5ml, add 0.05mol/L acetic acid-sodium-acetate buffer 1.5ml, one of filter paper bar of 1 * 6cm Xinhua, is hydrolyzed 60min at 55 ℃.After hydrolysis, adopt the glucose content in following DNS method mensuration system, and calculate the enzyme activity of cellulase.
In following enforcement, in fermention medium, the measuring method of glucose content adopts DNS method, specific as follows:
(1) preparation of DNS reagent: 200g Seignette salt, be dissolved in a certain amount of water, heating for dissolving, adds 10.0g3,5-dinitrosalicylic acid, 10g NaOH adds 2g phenol, 0.5g sodium sulphite anhydrous 99.3 after dissolving, all, after heating for dissolving, be cooled to room temperature, be settled to 1000ml.
(2) drafting of glucose typical curve: accurately take the analytically pure dextrose anhydrous of 100.0mg (being dried to constant weight at 105 ℃ in advance), after dissolving with a small amount of distilled water, quantitatively transfer in 100ml volumetric flask, then constant volume to scale, shake up, concentration is 1mg/mL.Get 10 test tubes, add respectively after glucose standardized solution 0.2mL, 0.4mL, 0.6mL, 0.8mL, 1.0mL, 1.2mL, 1.6mL, with distilled water diluting, to 2mL, then add 2.5mL DNS reagent mix even, in boiling water, heat 5min.After taking-up, with being water-cooled to room temperature, constant volume, to 25ml, shakes up.After 30min, under 520nm, survey OD value.With OD
520value is ordinate zou, and glucose content is X-coordinate drawing standard curve, and form is Y=aX+b.
(3) mensuration of glucose concn in fermented liquid: get and be diluted to certain density fermented supernatant fluid (4000r/min, centrifugal 20min), add 2.5mL DNS reagent mix even, heat 5min in boiling water.Take out water cool to room temperature, be settled to 25mL, shake up, after standing 30min, in 520nm, go out to survey OD
520value.OD per sample
520value is calculated the glucose content in fermented liquid with typical curve.
Separation and the evaluation of embodiment 1, Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101
One, the separation of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101
Tree peony (Paeonia ostii from Kingsoft village, Shun An town, Tongling Anhui root bark of tree peony planting site, Feng Dan) rhizosphere of (6 years ages of strain) plant gathers soil sample, soil sample is put into the little triangular flask that sterilized water and sterilizing granulated glass sphere are housed, the 200rpm/min 20min that vibrates; The suspension of getting after vibration is done serial gradient dilution, 10
3, 10
4extent of dilution suspension is respectively drawn 200 μ l and is evenly coated with
nitrogen-free agar, puts 30 ℃ and cultivates after 4d, picking list bacterium colony,
on nitrogen-free agar solid medium flat board, with plate streak, carry out purifying agaric.By one of them bacterial strain called after tree peony rhizosphere azotobacter CSM1101 of separation and purification gained.
Two, the evaluation of tree peony rhizosphere azotobacter CSM1101
1. morphological specificity is observed and physio-biochemical characteristics mensuration
Morphology and physio-biochemical characteristics are identified with reference to " uncle Jie Shi systematic bacteriology handbook " (Garrity, 2001) and " the conventional authentication method of general bacteriology " (eastern elegant pearl etc., 2001), according to ordinary method, are carried out.
The morphological feature of tree peony rhizosphere azotobacter CSM1101 is as follows: Gram-positive, and motion, it is shaft-like that thalline is, and diameter is 0.6-0.8 μ m, and long is 2.0-7.6 μ m.Gemma column, 1.6-3.5 * 1.2-1.5 μ m, middle life is raw to time end.Sporocyst obviously enlarges into spindle-type or bar-shaped.The irregular splintery of its colony edge, translucent to opaque.Rear its bacterium colony is coarse, shallow white, middle slightly projection.Bacterium colony adheres to media surface.
The physiological and biochemical property measurement result of tree peony rhizosphere azotobacter CSM1101 is as follows:
Hydrolyzed starch: feminine gender;
Caseinhydrolysate: feminine gender;
Gelatin hydrolysate: feminine gender;
Utilize Citrate trianion: feminine gender;
Tyrosine decomposes: feminine gender;
Phenylalanine deaminase experiment: feminine gender;
Yolk lecithin enzyme experiment: feminine gender;
Indoles produces: feminine gender;
Hippurate experiment: feminine gender;
Catalase experiment: the positive;
Nitrate reduction experiment: the positive;
Protosol produces: the positive;
Anti-N,O-Diacetylmuramidase experiment: the positive;
Decomposition glucose produces sour aerogenesis: the positive;
Decompose pectinose and produce sour aerogenesis: the positive;
Decompose wood sugar and produce sour aerogenesis: the positive;
Decompose N.F,USP MANNITOL and produce sour aerogenesis: the positive;
Salt tolerance test: 7%NaCl growth;
In the nutrient broth medium (NB) of pH6.8 and the Sharpe glucose broth of pH5.7, all can grow.On MR-VP meat soup, produce acetyl methyl carbinol, pH6.0.
The sequence amplification of 2.16S rDNA and analysis
Adopt bacterial genomes DNA extraction test kit (TIANGEN company) to extract genomic dna as the template of PCR reaction.PCR primer (Pf5 '-CGGGATCCAGAGTTTGATCCTGGCTCAG-3 ' and Pr5 '-CGGGATCCAAGGAGGTGATCCAGCC-3 ') synthetic by life work biotechnology (Shanghai) limited-liability company.Use the 16S rDNA of Perkin-ElmerGeneAmp PCR System9700PCR amplification bacterium.Reaction system is 10 * PCR buffer, 2.5uL; DNTP(2.5mM) (TaKaRa), 2uL, Pf(10pmol/uL) 0.1uL, Pr(10pmol/uL) 0.1uL, Taq polysaccharase (5U/uL) (TaKaRa), 0.125uL, genomic dna 0.5uL, adds ddH
2o to 25uL.PCR reaction conditions is, 94 ℃ of denaturation 5min, and 94 ℃ of sex change 1min, 52 ℃ of renaturation 1min, 72 ℃ are extended 1min, and 72 ℃ last extends 10min, 30 circulations.Pcr amplification product is by giving birth to work biotechnology (Shanghai) limited-liability company purifying order-checking.The 16S rDNA sequence of bacterial strain is carried out to sequence comparing analysis at GenBank GenBank.
Result shows the tree peony rhizosphere azotobacter CSM110116S rDNA sequence total length 1496bp recording, as SEQID No.1.The result of carrying out blastn analysis at GENBANK shows, its sequence and Paenibacillus polymyxa strain M-1(EF656457) similarity reach 99%.
According to the morphological specificity of tree peony rhizosphere azotobacter CSM1101, physiological and biochemical property and 16s rDNA sequence homology analysis result, tree peony rhizosphere azotobacter CSM1101 is accredited as to Paenibacillus polymyxa (Paenibacillus polymyxa).Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on December 09th, 2013 and (is called for short CGMCC, address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City), deposit number is CGMCC No.8527.
The biological activity of embodiment 2, Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101
One, the nitrogenase activity of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 is measured
Nitrogenase activity is measured the Acetylene Reduction method that adopts.In 15mm * 15mm screw-cap test tube, add 2.5mL semi-solid
nitrogen-free agar, after inoculation Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527, has filled in tampon, cultivates 24h for 30 ℃; Change again aseptic plug sealing, extract 10% gas out, inject the acetylene of same volume, cultivate 24h for 30 ℃.Extract gas sample and measure ARA(acetyiene reduction activity, acetyiene reduction activity on Varian VISTA6000 type gas chromatograph).Three repetitions are established in experiment, get its mean value and calculate variance to be not more than 5%.
Measurement result shows, CSM1101 exists
in nitrogen-free agar, shown higher nitrogenase activity.Its acetyiene reduction activity is 12.6C
2h
4nmol mL
-1h
-1.
Two, the bacteriostatic activity of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101
With tree peony botrytis (Botrytis paeoniae) ACCC36053, tree peony branch spore mould (Cladosporium paeoniae) ACCC36063, arbitrary bacterial strain in the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) is separately pathogenic bacteria, all adopt dull and stereotyped face-off method to measure bacteriostasis rate, concrete grammar is as follows: after pathogenic bacteria bacterial strain activates on PDA inclined-plane, add sterilized water to make pathogenic bacteria suspension, pathogenic bacteria suspension is added in the PDA substratum that is cooled to 45-50 ℃ after thawing and mixes and be down flat plate, at 30 ℃, cultivate 4 days, obtain pathogenic bacteria flat board, with the punch tool that diameter is 6mm, beat and get pathogenic bacteria bacterium cake.
Measure as follows the bacteriostasis rate of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 to every kind of pathogenic bacteria: experiment in triplicate, repeats pathogenic bacteria at every turn and establishes two processing, i.e. CSM1101 group and control group.20 aseptic PDA flat boards are divided into two groups, be CSM1101 group and control group, every group of 10 flat boards, carry out following inoculation processing: the pathogenic bacteria bacterium cake that is 6mm at the dull and stereotyped center inoculation of the aseptic PDA of CSM1101 group diameter, is inoculated in Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 slant activation bacterial classification apart from pathogenic bacteria bacterium cake 3cm place.Meanwhile, at the dull and stereotyped center of aseptic PDA of control group, only inoculate the pathogenic bacteria bacterium cake that diameter is 6mm.CSM1101 group flat board and the control group flat board through above-mentioned inoculation, processed are cultivated to 5-7 days at 30 ℃, when control group dull and stereotyped (only inoculating pathogenic bacteria) covers with whole culture dish, measure the pathogenic bacteria colony diameter of control group and the pathogenic bacteria colony diameter of CSM1101 group, calculate bacteriostasis rate.
Pathogenic bacteria colony diameter * 100 of bacteriostasis rate (%)=(the pathogenic bacteria colony diameter of the pathogenic bacteria colony diameter of control group-CSM1101 group)/control group.
Result is as shown in table 1, show that Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 all has stronger dull and stereotyped bacteriostatic activity to tree peony botrytis (Botrytis paeoniae) ACCC36053, mould (Cladosporium paeoniae) ACCC36063 of tree peony branch spore and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter), its bacteriostasis rate has reached respectively 57.2%, 65.7% and 88.3%.
The bacteriostasis rate of table 1 Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 to pathogenic bacteria
The raw material that following embodiment 3-6 adopts is all identical, and just processing condition are variant, as shown in table 2.
The processing condition of table 2, embodiment 3-6
Embodiment 3, utilize maize straw enzymolysis solution produce to promote the microbial inoculum A of tree peony growth antagonism pathogenic bacteria
1, the preparation of fermention medium
Get maize straw 8000g(with dry weight basis), be cut into segment, drop into hydrothermal reaction kettle, under the condition of 180 ℃ and 1.0Mpa, carry out naturally drying after hydrothermal treatment consists 20min, obtain through pretreated maize straw; By this after pretreated corn stalk powder is broken to 100 orders, according to adding the ratio of 15FPU to add cellulase with every gram of stalk of dry weight basis, according to adding the ratio of 1L distilled water to add distilled water with the every 200 grams of dry straws of dry weight basis, in 55 ℃, 100rpm(rotation radius 20mm), cooling centrifugal after the Water Under solution 48h of pH=4.8, collect respectively supernatant liquor and precipitation, this supernatant liquor is enzymolysis solution, and this precipitation is enzymolysis residue.Measure the glucose content in enzymolysis solution, result shows that the glucose content in enzymolysis solution is 55g/L.With enzymolysis solution, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water preparation fermention medium (is that fermention medium is by enzymolysis solution, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water forms), it is 20g that the content of enzymolysis solution described in every liter of described fermention medium meets the glucose content of every liter of described fermention medium; In every liter of described fermention medium, (NH
4)
2sO
4content be 10g, MgSO
4content be 0.4g, CaCO
3content be that the content of 6g, corn steep liquor is 2g.
2, fermentation culture
2.1 seed culture
Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 is got to 2-3 ring in NA medium slant after activation culture be inoculated in the 50ml NB substratum in 500ml triangular flask.28 ℃, 26mm amplitude, 24h is as seed liquor for 200r/min shaking culture, prepares to cultivate for ferment tank.
2.2 fermentation culture
The fermention medium of getting 50L step 1 is placed in 100L automatic fermenter, in 121 ℃ of sterilizing 20min, cooling after, by the seed liquor access fermention medium of 2.1 gained, inoculum size (V/V) is 10%.Culture temperature is 30 ℃, by the dissolved oxygen amount that regulates air flow and mixing speed to maintain in fermention medium, is 30%, and in fermenting process, automatically regulating pH is 6.8-7.2.While cultivating 14 hours, add the enzymolysis solution of step 1, making the glucose content in fermention medium is 15g/L, continues to cultivate 16 hours, finishes fermentation.Altogether ferment 30 hours.Get fermented liquid, measure the thalline content in fermented liquid, test in triplicate results averaged.Result shows that the content of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 of embodiment 1 in fermented liquid is 2.58 * 10
10cfu/mL, gemma content is 85%.By the mass mixings such as enzymolysis residue in this fermented liquid and step 1, after spraying is dry, (60 ℃) granulate, and are promoted the microbial inoculum A of tree peony growth antagonism pathogenic bacteria.
Embodiment 4, utilize maize straw enzymolysis solution produce to promote the microbial inoculum B of tree peony growth antagonism pathogenic bacteria
1, the preparation of fermention medium
Get maize straw 8000g(with dry weight basis), be cut into segment, drop into hydrothermal reaction kettle, under the condition of 180 ℃ and 1.0Mpa, carry out naturally drying after hydrothermal treatment consists 20min, obtain through pretreated maize straw; By this after pretreated corn stalk powder is broken to 100 orders, according to adding the ratio of 15FPU to add cellulase with every gram of stalk of dry weight basis, according to adding the ratio of 1L distilled water to add distilled water with the every 200 grams of dry straws of dry weight basis, in 55 ℃, 100rpm(rotation radius 20mm), cooling centrifugal after the Water Under solution 48h of pH=4.8, collect respectively supernatant liquor and precipitation, this supernatant liquor is enzymolysis solution, and this precipitation is enzymolysis residue.Measure the glucose content in enzymolysis solution, result shows that the glucose content in enzymolysis solution is 55g/L.With enzymolysis solution, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water preparation fermention medium (is that fermention medium is by enzymolysis solution, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water forms), it is 20g that the content of enzymolysis solution described in every liter of described fermention medium meets the glucose content of every liter of described fermention medium; In every liter of described fermention medium, (NH
4)
2sO
4content be 10g, MgSO
4content be 0.4g, CaCO
3content be that the content of 6g, corn steep liquor is 2g.
2, fermentation culture
2.1 seed culture
Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 is got to 2-3 ring in NA medium slant after activation culture be inoculated in the 50ml NB substratum in 500ml triangular flask.28 ℃, 26mm amplitude, 24h is as seed liquor for 200r/min shaking culture, prepares to cultivate for ferment tank.
2.2 fermentation culture
The fermention medium of getting 50L step 1 is placed in 100L automatic fermenter, in 121 ℃ of sterilizing 20min, cooling after, by the seed liquor access fermention medium of 2.1 gained, inoculum size (V/V) is 10%.Culture temperature is 30 ℃, by the dissolved oxygen amount that regulates air flow and mixing speed to maintain in fermention medium, is 30%, and in fermenting process, automatically regulating pH is 6.8-7.2.Cultivate 24 hours, finish fermentation.Get fermented liquid, measure the thalline content in fermented liquid, test in triplicate results averaged.Result shows that the content of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 of embodiment 1 in fermented liquid is 1.86 * 10
8cfu/mL, gemma content is 85%.By the mass mixings such as enzymolysis residue in fermented liquid and step 1, after spraying is dry, (60 ℃) granulate, and are promoted the microbial inoculum B of tree peony growth antagonism pathogenic bacteria.
Embodiment 5, utilize maize straw enzymolysis solution produce to promote the microbial inoculum C of tree peony growth antagonism pathogenic bacteria
1, the preparation of fermention medium
Get maize straw 8000g(with dry weight basis), be cut into segment, drop into hydrothermal reaction kettle, under the condition of 200 ℃ and 2.0Mpa, carry out naturally drying after hydrothermal treatment consists 20min, obtain through pretreated maize straw; By this after pretreated corn stalk powder is broken to 100 orders, according to adding the ratio of 15FPU to add cellulase with every gram of stalk of dry weight basis, according to adding the ratio of 1L distilled water to add distilled water with the every 200 grams of dry straws of dry weight basis, in 55 ℃, 100rpm(rotation radius 20mm), cooling centrifugal after the Water Under solution 48h of pH=4.8, collect respectively supernatant liquor and precipitation, this supernatant liquor is enzymolysis solution, and this precipitation is enzymolysis residue.Measure the glucose content in enzymolysis solution, result shows that the glucose content in enzymolysis solution is 60g/L.With enzymolysis solution, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water preparation fermention medium (is that fermention medium is by enzymolysis solution, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water forms), it is 20g that the content of enzymolysis solution described in every liter of described fermention medium meets the glucose content of every liter of described fermention medium; In every liter of described fermention medium, (NH
4)
2sO
4content be 10g, MgSO
4content be 0.4g, CaCO
3content be that the content of 6g, corn steep liquor is 2g.
2, fermentation culture
2.1 seed culture
Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 is got to 2-3 ring in NA medium slant after activation culture be inoculated in the 50ml NB substratum in 500ml triangular flask.28 ℃, 26mm amplitude, 24h is as seed liquor for 200r/min shaking culture, prepares to cultivate for ferment tank.
2.2 fermentation culture
The fermention medium of getting 50L step 1 is placed in 100L automatic fermenter, in 121 ℃ of sterilizing 20min, cooling after, by the seed liquor access fermention medium of 2.1 gained, inoculum size (V/V) is 10%.Culture temperature is 30 ℃, by the dissolved oxygen amount that regulates air flow and mixing speed to maintain in fermention medium, is 30%, and in fermenting process, automatically regulating pH is 6.8-7.2.Cultivate 24 hours, finish fermentation.Get fermented liquid, measure the thalline content in fermented liquid, test in triplicate results averaged.Result shows that the content of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 of embodiment 1 in fermented liquid is 2.78 * 10
9cfu/mL, gemma content is 85%.By the mass mixings such as enzymolysis residue in fermented liquid and step 1, after spraying is dry, (60 ℃) granulate, and are promoted the microbial inoculum C of tree peony growth antagonism pathogenic bacteria.
Embodiment 6, utilize maize straw enzymolysis solution produce to promote the microbial inoculum D of tree peony growth antagonism pathogenic bacteria
1, the preparation of fermention medium
Get maize straw 8000g(with dry weight basis), be cut into segment, drop into hydrothermal reaction kettle, under the condition of 200 ℃ and 2.0Mpa, carry out naturally drying after hydrothermal treatment consists 10min, obtain through pretreated maize straw; By this after pretreated corn stalk powder is broken to 100 orders, according to adding the ratio of 15FPU to add cellulase with every gram of stalk of dry weight basis, according to adding the ratio of 1L distilled water to add distilled water with the every 200 grams of dry straws of dry weight basis, in 55 ℃, 100rpm(rotation radius 20mm), cooling centrifugal after the Water Under solution 48h of pH=4.8, collect respectively supernatant liquor and precipitation, this supernatant liquor is enzymolysis solution, and this precipitation is enzymolysis residue.Measure the glucose content in enzymolysis solution, result shows that the glucose content in enzymolysis solution is 46g/L.With enzymolysis solution, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water preparation fermention medium (is that fermention medium is by enzymolysis solution, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water forms), it is 20g that the content of enzymolysis solution described in every liter of described fermention medium meets the glucose content of every liter of described fermention medium; In every liter of described fermention medium, (NH
4)
2sO
4content be 10g, MgSO
4content be 0.4g, CaCO
3content be that the content of 6g, corn steep liquor is 2g.
2, fermentation culture
2.1 seed culture
Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 is got to 2-3 ring in NA medium slant after activation culture be inoculated in the 50ml NB substratum in 500ml triangular flask.28 ℃, 26mm amplitude, 24h is as seed liquor for 200r/min shaking culture, prepares to cultivate for ferment tank.
2.2 fermentation culture
The fermention medium of getting 50L step 1 is placed in 100L automatic fermenter, in 121 ℃ of sterilizing 20min, cooling after, by the seed liquor access fermention medium of 2.1 gained, inoculum size (V/V) is 10%.Culture temperature is 30 ℃, by the dissolved oxygen amount that regulates air flow and mixing speed to maintain in fermention medium, is 30%, and in fermenting process, automatically regulating pH is 6.8-7.2.Cultivate 24 hours, finish fermentation.Get fermented liquid, measure the thalline content in fermented liquid, test in triplicate results averaged.Result shows that the content of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 of embodiment 1 in fermented liquid is 4.42 * 10
8cfu/mL, gemma content is 85%.By the mass mixings such as enzymolysis residue in fermented liquid and step 1, after spraying is dry, (60 ℃) granulate, and are promoted the microbial inoculum D of tree peony growth antagonism pathogenic bacteria.
Comparative example: utilize starchy material to produce and promote the also microbial inoculum E of antagonism pathogenic bacteria of tree peony growth
1, fermention medium preparation
Fermention medium: with starch, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water preparation fermention medium (is that fermention medium is by starch, (NH
4)
2sO
4, MgSO
4, CaCO
3, corn steep liquor and water forms), it is 20g that the content of starch described in every liter of described fermention medium meets the glucose content of every liter of described fermention medium; In every liter of described fermention medium, (NH
4)
2sO
4content be 10g, MgSO
4content be 0.4g, CaCO
3content be that the content of 6g, corn steep liquor is 2g.）
The component adopting in the fermention medium of this comparative example is except the starch of the glucose contents such as maize straw enzymolysis solution is replaced with, and other is all identical with embodiment 3-6 for other component in fermention medium.
2, fermentation culture
In this fermentation culture, the preparation method of fermented liquid is identical with embodiment 4-6, specific as follows:
2.1 seed culture
Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 is got to 2-3 ring in NA medium slant after activation culture be inoculated in the 50ml NB substratum in 500ml triangular flask.28 ℃, 26mm amplitude, 24h is as seed liquor for 200r/min shaking culture, prepares to cultivate for ferment tank.
2.2 fermentation culture
Get 50L fermention medium and be placed in 100L automatic fermenter, in 121 ℃ of sterilizing 20min, cooling after, by the seed liquor access fermention medium of 2.1 gained, inoculum size (V/V) is 10%.Culture temperature is 30 ℃, by the dissolved oxygen amount that regulates air flow and mixing speed to maintain in fermention medium, is 30%, and in fermenting process, automatically regulating pH is 6.8-7.2.Cultivate 24 hours, finish fermentation.Get fermented liquid, measure the thalline content in fermented liquid, test in triplicate results averaged.Result shows that the content of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 of embodiment 1 in fermented liquid is 3.66 * 10
9cfu/mL, gemma content is 85%.(60 ℃) after fermented liquid spray drying are granulated, be promoted the microbial inoculum E of tree peony growth antagonism pathogenic bacteria.
The microbial inoculum of embodiment 7, the growth of promotion tree peony antagonism pathogenic bacteria promotes Paeonia suffruticosa seed to sprout and tree peony growth of seedling
Seed collection: take pericarp crab oil July as standard Feng Dan (Paeonia ostii) the Follicle radish fruit (Kingsoft village, Shun An town, Tongling Anhui) of gathering, put indoor drying in the shade and collect seed, 0.5%KMnO to pericarp cracking
4feng Dan (Paeonia ostii) seed that immersion 2h carries out disinfection after being sterilized is for test.Test site is Shanghai Songjiang district Shanghai Chenshan Botanical Garden.
The preparation of Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 bacteria suspension: the microbial inoculum A of the promotion tree peony growth of embodiment 3 antagonism pathogenic bacteria is suspended in sterilized water and obtains Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 bacteria suspension, and making the content of Paenibacillus polymyxa in bacteria suspension (Paenibacillus polymyxa) CSM1101CGMCC No.8527 is 10
9cfu/mL.
Cancel Feng Dan (Paeonia ostii) seed after poison, Feng Dan (Paeonia ostii) seed after being soaked seed with 50 ℃ of sterilized water seed soaking 24h, hereinafter to be referred as the Paeonia suffruticosa seed after seed soaking, carry out take root (radicle sproutings) experiment and germinate (the plumule sprouting) of following seed and test.Take root (radicle sprouting) experiment and (plumule sproutings) experiment of germinateing of seed all in triplicate, repeats to arrange two kinds of processing at every turn, and microbial inoculum group lamination is processed and the processing of control group lamination.Seed is taken root in (radicle sprouting) experiment, every kind of Paeonia suffruticosa seed of processing after 300 seed soaking.Germinate in (plumule sprouting) experiment, 50 main root length >=40mm and the consistent seed of taking root of length are chosen in every kind of processing.Specific experiment method is as follows:
1, seed take root (radicle sprouting) experiment
Microbial inoculum group lamination is processed with control group lamination and is processed in these two kinds of processing except the fore-pretreatment method of taking root is different, and other processing mode is identical.Microbial inoculum group lamination is processed take root before pre-treatment as follows: the Paeonia suffruticosa seed after seed soaking is placed in to Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 bacteria suspension (10
9cfu/mL) in, at 25-30 ℃, soak 1 hour, complete take root before pre-treatment.Control group lamination is processed take root before pre-treatment as follows: the Paeonia suffruticosa seed after seed soaking is placed in to sterilized water at 25-30 ℃ and soaks 1 hour, complete take root before pre-treatment.Then will complete pretreated seed before taking root and all carry out following root culture: will complete before taking root after the perlite (lamination matrix) of pretreated seed and the water content high-temperature sterilization that is 400% mixes according to the volume ratio of 1:3, packing polyethylene plastic bag into is all placed in 10 ℃-25 ℃ (daytimes 20-25 ℃, night 10-15 ℃) and carries out root culture.Observe investigation rootage duration, the seed percentage of root culture 90d researching determining rooting rate, main root length, main root length >=40mm every day 1 time.Data are carried out significance of difference analysis with SPSS software.
Wherein, take radicle to expose the length of kind of skin be seed length 1/2 for taking root.
Rooting rate=(L1/L0) * 100%;
L1: the seed grain number of taking root in the experiment seed of taking root; L0: the experiment seed grain number of taking root.
2, germination (plumule sprouting) experiment
The microbial inoculum group lamination of getting respectively step 1 is processed and control group lamination is processed main root length >=40mm and the consistent seed of taking root of length that these two kinds of processing obtain and preserved after 21 days at 4 ℃, plant and be equipped with in peat and perlitic cave dish in high 10cm, be placed in 10-25 ℃ of greenhouse (daytime 20-25 ℃, night 10-15 ℃) in germinate and cultivate seedling, observe every day 1 time, measure germinating time, germinate and cultivate the dry weight that 90d measures percentage of germination, height of seedling, whole tree peony seedling plant, calculate percentage of germination.Data are carried out significance of difference analysis with SPSS software.
Wherein, take plumule exposes kind of skin as germination.
Percentage of germination=(L1 '/L0 ') * 100%;
L1 ': chitting piece grain number in Seed germination seed; L0 ': Seed germination seed grain number.
Result is as shown in table 3-table 5, and the rootage duration that microbial inoculum group lamination is processed is processed and shifted to an earlier date 10 days than control group lamination; The rooting rate that microbial inoculum group lamination is processed reaches 95.0%, and the rooting rate of processing than control group lamination has improved 33.8%; The main root length that microbial inoculum group lamination is processed is 69.0mm, is 1.68 times that control group lamination is processed; The seed percentage of main root length >=40mm that microbial inoculum group lamination is processed reaches 90 ± 0.5%, is 1.3 times that control group lamination is processed; The germinating time that microbial inoculum group lamination is processed is processed and has been shortened 10 days than control group lamination; The percentage of germination that microbial inoculum group lamination is processed reaches 99.0%, is 1.2 times that control group lamination is processed; The height of seedling that microbial inoculum group lamination is processed is 1.77 times that control group lamination is processed, and the dry weight that microbial inoculum group lamination is processed is 2.27 times that control group lamination is processed.Explanation be take the microbial inoculum of promotion tree peony growth that Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101CGMCC No.8527 is activeconstituents antagonism pathogenic bacteria and has been promoted significantly taking root (radicle sprouting) of Paeonia suffruticosa seed and germinateed (plumule sprouting) and the growth of tree peony seedling.Fig. 1 has shown the processing of microbial inoculum group lamination and the control group lamination treating part seed root culture situation of 60 days.
Table 3, microbial inoculum are processed the impact that Paeonia suffruticosa seed is taken root
Note: the different significance degree of the English alphabet differential of same column in table, between the processing of same letter in 0.05 level without significant difference, between the different processing of letter, in 0.05 level, there were significant differences.Rootage duration refers to the 1st time that seed is taken root.
Table 4, microbial inoculum are processed the impact that Paeonia suffruticosa seed is germinateed
| Process | Germinating time (my god) | Percentage of germination (%) |
| Microbial inoculum group lamination is processed | 26±1a | 99.0±0.6a |
| Control group lamination is processed | 36±2b | 81.0±0.2b |
Note: the different significance degree of the English alphabet differential of same column in table, between the processing of same letter in 0.05 level without significant difference, between the different processing of letter, in 0.05 level, there were significant differences.Germinating time refers to the time of the seed germination that takes root of the 1st main root length >=40mm.
Table 5, microbial inoculum are processed the impact on tree peony growth of seedling
| Process | Height of seedling (mm) | Dry weight (mg) |
| Microbial inoculum group lamination is processed | 122.0±2.0a | 501.0±5.0a |
| Control group lamination is processed | 69.0±2.0b | 221.0±3.0b |
Note: the different significance degree of the English alphabet differential of same column in table, between the processing of same letter in 0.05 level without significant difference, between the different processing of letter, in 0.05 level, there were significant differences.Height of seedling is the height of seedling over-ground part, and dry weight is the dry weight of whole plant, comprises root and bud.
Claims (10)
1. Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101, the deposit number at Qi China Committee for Culture Collection of Microorganisms common micro-organisms center is CGMCC No.8527.
2. a microbial inoculum, its activeconstituents is Paenibacillus polymyxa claimed in claim 1 (Paenibacillus polymyxa) CSM1101.
3. microbial inoculum according to claim 2, is characterized in that: described microbial inoculum is following A 1)-A3) in any microbial inoculum:
A1) promote the microbial inoculum of tree peony growth and antagonism pathogenic bacteria;
Described pathogenic bacteria be in tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) three kinds, any two or any;
A2) microbial inoculum of antagonism pathogenic bacteria; Described pathogenic bacteria be in tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) three kinds, any two or any;
A3) promote the microbial inoculum of tree peony growth.
Paenibacillus polymyxa claimed in claim 1 (Paenibacillus polymyxa) CSM1101 preparation following A 1)-A3) and in any microbial inoculum in application:
A1) promote the microbial inoculum of tree peony growth and antagonism pathogenic bacteria;
Described pathogenic bacteria be in tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) three kinds, any two or any;
A2) microbial inoculum of antagonism pathogenic bacteria; Described pathogenic bacteria be in tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) three kinds, any two or any;
A3) promote the microbial inoculum of tree peony growth.
5. microbial inoculum according to claim 3 or application claimed in claim 4, is characterized in that: the growth of described promotion tree peony is presented as following B1)-B7) in any:
B1) shorten Paeonia suffruticosa seed rootage duration;
B2) improve Paeonia suffruticosa seed rooting rate;
B3) shorten the germinating time of Paeonia suffruticosa seed;
B4) improve the percentage of germination of Paeonia suffruticosa seed;
B5) improve the main root length of tree peony;
B6) improve tree peony height of seedling;
B7) improve tree peony Seedling Biomass.
6. the purposes of the microbial inoculum described in Paenibacillus polymyxa claimed in claim 1 (Paenibacillus polymyxa) CSM1101 or claim 2,3 or 5, described purposes is following C1)-C3) in any:
C1) promote tree peony growth and antagonism pathogenic bacteria;
C2) described promotion tree peony growth;
C3) described antagonism pathogenic bacteria.
7. promote the method for tree peony growth, be included in before taking root the Paeonia suffruticosa seed after soaking seed is processed to the step of carrying out again root culture for 1-3 hour with the microbial inoculum described in claim 2 or 3.
8. purposes according to claim 6 or method claimed in claim 7, is characterized in that: the growth of described promotion tree peony is presented as following B1)-B7) in any:
B1) shorten Paeonia suffruticosa seed rootage duration;
B2) improve Paeonia suffruticosa seed rooting rate;
B3) shortening the first of Paeonia suffruticosa seed sprouts the phase;
B4) improve the percentage of germination of Paeonia suffruticosa seed;
B5) improve the main root length of tree peony;
B6) improve tree peony height of seedling;
B7) improve tree peony Seedling Biomass;
Described antagonism pathogenic bacteria be in antagonism tree peony botrytis (Botrytis paeoniae), tree peony branch spore mould (Cladosporium paeoniae) and the variegated tail spore of Chinese herbaceous peony mould (Cercospora varricolor Winter) three kinds, any two or any.
9. the method for Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 described in cultivation claim 1, comprises Paenibacillus polymyxa (Paenibacillus polymyxa) CSM1101 in the step of cultivating for cultivating the substratum of Paenibacillus polymyxa.
10. the preparation method of microbial inoculum described in claim 2,3 or 5, comprises the steps: Paenibacillus polymyxa claimed in claim 1 (Paenibacillus polymyxa) CSM1101 as activeconstituents, to obtain described microbial inoculum.
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| CN201610308942.5A CN105861378B (en) | 2014-03-26 | 2014-03-26 | The application of the Paenibacillus polymyxa of a variety of tree peony pathogens of antagonism |
| CN201410117673.5A CN104073451B (en) | 2014-03-26 | 2014-03-26 | Paenibacillus polymyxa and the application thereof of the multiple tree peony pathogen of one strain antagonism |
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| CN201410117673.5A Expired - Fee Related CN104073451B (en) | 2014-03-26 | 2014-03-26 | Paenibacillus polymyxa and the application thereof of the multiple tree peony pathogen of one strain antagonism |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105053015A (en) * | 2015-07-15 | 2015-11-18 | 河南科技大学 | Biopesticide for controlling cercospora varrireolorWinter, and preparation method for biopesticide |
| CN105907657A (en) * | 2016-03-25 | 2016-08-31 | 洛阳理工学院 | Peony endogenous Bacillus amyloliquefaciens and its separation method and use |
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| CN1169942A (en) * | 1996-02-15 | 1998-01-14 | 莱雅公司 | Container for separate packaging, then mixing, of at least two products |
| CN1169942C (en) * | 2002-12-04 | 2004-10-06 | 上海泽元海洋生物技术有限公司 | Microbe preparation for curing bacteroidal grancille wilt of plant and its method as well as usage |
| US20130014434A1 (en) * | 2011-07-12 | 2013-01-17 | Tegraseal Products, Llc | Compositie delivery and nutrient system for agricultural and nursery uses |
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2014
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Patent Citations (3)
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| CN1169942A (en) * | 1996-02-15 | 1998-01-14 | 莱雅公司 | Container for separate packaging, then mixing, of at least two products |
| CN1169942C (en) * | 2002-12-04 | 2004-10-06 | 上海泽元海洋生物技术有限公司 | Microbe preparation for curing bacteroidal grancille wilt of plant and its method as well as usage |
| US20130014434A1 (en) * | 2011-07-12 | 2013-01-17 | Tegraseal Products, Llc | Compositie delivery and nutrient system for agricultural and nursery uses |
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| 苍桂璐等: "多粘类芽孢杆菌的研究进展", 《安徽农业科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105053015A (en) * | 2015-07-15 | 2015-11-18 | 河南科技大学 | Biopesticide for controlling cercospora varrireolorWinter, and preparation method for biopesticide |
| CN105053015B (en) * | 2015-07-15 | 2017-10-27 | 河南科技大学 | It is a kind of to be used to prevent and treat biological pesticide of tree peony brown spot and preparation method thereof |
| CN105907657A (en) * | 2016-03-25 | 2016-08-31 | 洛阳理工学院 | Peony endogenous Bacillus amyloliquefaciens and its separation method and use |
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| CN105861378B (en) | 2019-06-25 |
| CN105861378A (en) | 2016-08-17 |
| CN104073451B (en) | 2016-05-18 |
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