CN106047965A - Preparation for preventing or treating eratocystis paradoxa and preparation method of preparation - Google Patents
Preparation for preventing or treating eratocystis paradoxa and preparation method of preparation Download PDFInfo
- Publication number
- CN106047965A CN106047965A CN201610424369.4A CN201610424369A CN106047965A CN 106047965 A CN106047965 A CN 106047965A CN 201610424369 A CN201610424369 A CN 201610424369A CN 106047965 A CN106047965 A CN 106047965A
- Authority
- CN
- China
- Prior art keywords
- preparation
- paenibacillus polymyxa
- bacteria suspension
- precipitation
- supernatant
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION 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
- A01N63/00—Biocides, 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
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/04—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using bacteria
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
Abstract
The invention provides a preparation for preventing or treating eratocystis paradoxa and a preparation method of the preparation. Detailed invention points are as follows: bacterial suspension of Paenibacillus polymyxa strain DSM 36 or an extract of the metabolite of the Paenibacillus polymyxa strain DSM 36 can be used for preventing or treating the eratocystis paradoxa. The strain has been deposited at German Collection Center of Microorganisms under access number of DSM 36 by the skilled in the art before the application date.
Description
Technical field
The invention belongs to biological technical field, be specifically related to a kind of prevention or treatment Caulis Sacchari sinensis pineapple disease biological preparation and
Its preparation method.
Background technology
Caulis Sacchari sinensis pineapple disease is a kind of fungal disease occurred the cane seedling phase, and Caulis Sacchari sinensis pineapple disease most distinct feature is that sugarcane kind
Susceptible initial stage otch two ends redden, and with the fragrance of Fructus Ananadis comosi, sick sugarcane interior tissue reddens therewith.Caulis Sacchari sinensis pineapple disease is sick
Former bacterium is unusual thielaviopsis sp.Sugarcane kind once be will result in kind of a stem by pathogenic bacterial infection and is injured and can not germinate, and germination rate reduces, and gives
Sugarcane production causes loss in various degree.
At present, domestic Caulis Sacchari sinensis pineapple disease based on chemicals preventing and treating, such as, produces upper often use carbendazim wettable powder
1000 times of diluents of agent soak sugarcane kind to be prevented and treated.Wherein commonly use powder and also have Metalaxyl-M CGA-173506 Diacloden FS, SC
Difenoconazole Fluoxastrobin, A meter Miao receives suspending agent, seed soaking spirit, and lime water is quickly soaked seed, 70% thiophanate methyl wettability
Powder, 50% benzene Lai Te, the proud miaow of benzene No. 44 (carbendazim), Rabcides, five oxygen fens crash.For many years, Caulis Sacchari sinensis pineapple disease preventing and treating
Chemicals is less, predominantly chemical agent.But excessively use chemicals inevitably to cause drug residue, pathogen
The problem such as drug resistance and environmental pollution.
In recent years, generally believe that Biological control is to solve one of most promising approach of farm crop fungus disease in the world,
So in the urgent need to inventing a kind of microorganism formulation that can effectively prevent and treat Caulis Sacchari sinensis pineapple disease.
Summary of the invention
The goal of the invention of the present invention is to provide a kind of microorganism formulation that can effectively prevent and treat Caulis Sacchari sinensis pineapple disease, can keep away
Exempt from excessively to use chemicals to cause the problem such as pesticide residues and pathogenic bacteria resistance to drugs increase.Concrete technical scheme is:
Select Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) after cultivating
The extract of bacteria suspension or this bacterium metabolite makes preparation as effective ingredient, is directly sprayed to need prevention or treatment
The Caulis Sacchari sinensis surface of Caulis Sacchari sinensis pineapple disease, plays prevention or therapeutic purposes.
Wherein, Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36), for existing micro-life
Thing, was preserved in Germany's Organism Depositary by those skilled in the art before the application day, and preserving number is DSM 36.
Those skilled in the art can contact this preservation mechanism and obtain this strain, telephone number :+49 (0) 531-2616-0, fax :+49
(0) 531-2616-418, mailbox: E-mail:contact@dsmz.de；Other matters, can arrive its official website https: //
Www.dsmz.de/contact.htmlLeibniz-Institut inquires about.
The extraction of above-mentioned preparation and preparation method are to take after Paenibacillus polymyxa bacteria suspension after cultivating is centrifuged to take
Its supernatant is as microorganism formulation；
Above-mentioned preparation preferably extracts and preparation method is:
1) taking bacteria suspension 1 parts by volume of Paenibacillus polymyxa, its effective bacteria concentration is 103-108Cfu/ml, 10000-
14000rpm takes supernatant after being centrifuged 8-12min；
2) in centrifuged supernatant, ethanol is added so that determining alcohol is 20%-90%, centrifugal after standing, take its precipitation, will
Precipitation is dissolved in the sterilized water of taken 2-6 times of parts by volume of bacteria suspension, both a kind of for preventing or treating Caulis Sacchari sinensis pineapple disease
Preparation.
Above-mentioned preferred extraction and preparation method be:
1) taking bacteria suspension 1 parts by volume of Paenibacillus polymyxa, its effective bacteria concentration is 106-107Cfu/ml, 12000rpm
Supernatant is taken after centrifugal 10min；
2) in centrifuged supernatant, ethanol is added so that determining alcohol is 70%, 5000rpm after standing 2 hours in 4 degrees Celsius
Centrifugal 10min, takes its precipitation, precipitation is dissolved in the sterilized water of taken 4 times of parts by volume of bacteria suspension, to obtain final product.
Another inventive point of the present invention is to provide a kind of system of preparation for preventing or treat Caulis Sacchari sinensis pineapple disease
Preparation Method, described preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in
Culture medium is cultivated, obtains bacteria suspension；
2) above-mentioned bacteria suspension takes supernatant after centrifugal treating, both obtains.
Wherein, preferred preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in
In LB fluid medium 30-35 degree Celsius, 200-240rpm concussion is cultivated 24-48 hour, and obtaining effective bacteria concentration is 103-
108The bacteria suspension of cfu/ml, wherein effectively bacteria concentration refers to the bacteria concentration in fermentation liquid；
2) take above-mentioned bacteria suspension after centrifugal treating, obtain supernatant, supernatant adds ethanol so that determining alcohol
For 20%-90%, it is centrifuged after standing, takes its precipitation, precipitation is dissolved in sterilized water, be prevention or treatment Caulis Sacchari sinensis pineapple disease
Preparation.
Wherein step 1, preferred method is: take Paenibacillus polymyxa (Paenibacillus polymyxa strain
DSM 36) strain be inoculated in LB fluid medium 33 degrees Celsius, 220rpm concussion is cultivated 36 hours, obtains effective bacteria concentration
It is 106-107The bacteria suspension of cfu/ml.
Wherein step 2, preferred method is: take the bacteria suspension obtained in 1 parts by volume step 1,10000-14000rpm from
After heart 8-12min, take supernatant and add ethanol so that determining alcohol is 60-80%, 2000-after stand at low temperature 1-3 hour
7000rpm is centrifuged 5-15min, takes its precipitation, precipitation is dissolved in the sterilized water of taken bacteria suspension parts by volume 2-6 times amount, obtains
The extract of Paenibacillus polymyxa metabolite, is prevention or the preparation for the treatment of Caulis Sacchari sinensis pineapple disease.
Wherein step 2, preferred method is: take the bacteria suspension obtained in 1 parts by volume step 1, and 12000rpm is centrifuged
After 10min, taking supernatant and add ethanol so that determining alcohol is 70%, after standing 2 hours in 4 degrees Celsius, 5000rpm is centrifuged
10min, takes its precipitation, precipitation is dissolved in the sterilized water of taken bacteria suspension parts by volume 4 times amount, is prevention or treatment Caulis Sacchari sinensis
The preparation of pineapple disease.
The above-mentioned training for cultivating Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36)
Foster base is LB fluid medium.
Mentioned microorganism preparation, has significantly prevention or therapeutical effect, by it to Caulis Sacchari sinensis Fructus Ananadis comosi to Caulis Sacchari sinensis pineapple disease
Disease pathogen bacterium unusual thielaviopsis sp inhibition is notable, and the sprouting to cane seedling simultaneously has obvious facilitation.
Paenibacillus polymyxa of the present invention (Paenibacillus polymyxa strain DSM 36) be from
In nature, screening and separating obtains, and through the comparison with existing strain, confirms and Germany's Organism Depositary, and preserving number is
The strain (Paenibacillus polymyxa strain DSM 36) of DSM 36 belongs to same strain.
For obtaining the microorganism formulation that can effectively prevent and treat Caulis Sacchari sinensis pineapple disease, the present invention has carried out as follows in research process
Experiment:
One, the discovery of effective microbe and separation
1, gather soil sample from river bank, Longjiang, Yizhou City bank, utilize Concentraton gradient isolation by dilution method to sieve bacterium: to weigh soil sample 10g,
Be equipped with in the conical flask of 90ml sterilized water prepare soil sample suspension, shake 30min, from suspension draw 10ml move into 90ml without
In the conical flask of bacterium water, till being diluted to 6 conical flasks successively, obtain the dilute of 10-2,10-3,10-4,10-5,10-6,10-7
Release liquid.Take respectively the diluent of 100ul10-4,10-5,10-6,10-7 coat LB culture medium (peptone 10g, yeast powder 5g,
NaCl 10g, agar powder 20g, distilled water 1000ml, pH 7.2～7.4) in, after being placed in 30 degree of incubators cultivation 48h, picking
Single bacterium colony that morphological characteristic is different, as strains tested after culture purified, experimentation is divided into and separates out 126 single strains.
2, Caulis Sacchari sinensis pineapple disease pathogen is inoculated into Martin fluid medium (KH2PO41g, MgSO 7H2O40.5g, egg
White peptone 5g, glucose 10g, distilled water 1000mL, pH7.2～7.4) in, 180rpm cultivates 48h and obtains indicator bacteria suspension, preparation
Martin solid (KH2PO41g, MgSO 7H2O40.5g, peptone 5g, glucose 10g, agar powder 20g, distilled water 1000mL,
PH7.2～7.4) culture plate coating indicator bacteria, it is put into just system for taking examination bacterium flat board for examination bacterium (truffle) from cultured
On the indicator bacteria flat board got ready, cultivate 3 days in 28 DEG C of constant incubators, observe for examination bacterium (truffle) around with or without inhibition zone and
Inhibition zone size.126 single strains of isolated in step 1, have all carried out 3 tests and have repeated screening, finally locked one
Strain aimed strain, its bacteriostatic activity in incubation is stable, and Antibacterial Activity is the strongest, and this bacterial strain inhibition zone reaches 27.8mm.
3, Main Morphology and the biological property of the aimed strain of said method isolated is: cell is shaft-like, leather orchid
Albert'stain Albert is positive (referring to accompanying drawing 1), has spore, and without soluble pigment on Nutrient agar, bacterium colony is thin and viscous, and surface is unglazed
Pool, translucent, growth optimum temperature is 33 DEG C, and growth optimal pH is 7.2.
4, aimed strain is identified: above-mentioned aimed strain is compared with ncbi database sequence after 16SrDNA checks order, knot
Fruit and Paenibacillus polymyxa strain DSM 36 (this bacterial strain is preserved in Germany's Organism Depositary at present,
Preserving number is DSM 36) homology, belong to same strain, therefore the bacterial strain sifted out is Paenibacillus polymyxa (Paenibacillus
polymyxa strain DSM 36)。
Two, Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is sick to Caulis Sacchari sinensis pineapple disease
The antagonistic activity of former bacterium
This test is with Caulis Sacchari sinensis pineapple disease pathogen as indicator bacteria, from cultured Paenibacillus polymyxa for examination bacterium flat board
Take and be put on the indicator bacteria flat board prepared (Caulis Sacchari sinensis pineapple disease pathogen) for examination truffle, be just placed in 28 DEG C of constant incubators
Cultivate 2 days, its antagonistic results such as Fig. 2 to pineapple disease pathogen
Three, Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) bacteriostatic active ingredients grinds
Study carefully
Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM is measured respectively by cup-plate method
36) bacteriostatic activity of bacteria suspension, centrifuged supernatant, bacteria-free filtrate, cell breakage liquid and heat treatment supernatant totally 5 kinds of solution,
Every kind of solution sets three parallel tests, and the concrete preparation method of described 5 kinds of solution is as follows:
(1) prepared by bacteria suspension: take Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM
36), during truffle is inoculated in LB liquid medium, 36h is cultivated in 33 DEG C of 220rpm concussions, obtains bacteria suspension (effective bacteria concentration 106-
107cfu/ml)；
(2) prepared by centrifuged supernatant: bacteria suspension 12000rpm takes supernatant after being centrifuged 10min；
(3) prepared by bacteria-free filtrate: takes the supernatant after above-mentioned steps 2 is centrifuged, obtains aseptic with 0.22um filtering with microporous membrane
Filtrate；
(4) cell breakage liquid: take the bacterial sediment after above-mentioned steps 2 is centrifuged and add appropriate amounts of sterilized water ice bath, ultrasonication
Cell obtains somatic cells and crushes liquid；
(5) heat treatment supernatant: take the supernatant after above-mentioned steps 2 is centrifuged and be sub-packed in test tube, 100 DEG C of water-bath 1h；
After above-mentioned 5 parts of test samples are disposed, prepare the culture plate containing indicator bacteria (Caulis Sacchari sinensis pineapple disease pathogen),
Each culture dish is placed 3 Oxford cups, each Oxford cup is separately added into bacteria suspension that the above-mentioned experimental procedure of 30ul obtained,
Centrifuged supernatant, bacteria-free filtrate, cell breakage liquid and heat treatment supernatant, cultivate 48h under the conditions of 28 DEG C, observe inhibition zone
The size that has that it's too late.
Table 1 Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) different disposal liquid is to sweet
The antagonistic activity of sugarcane pineapple disease pathogen
The above results shows: the fungistatic effect of bacteria suspension and centrifuged supernatant is suitable, and bacteria-free filtrate and somatic cells break
Broken liquid is without bacteriostatic activity, thus shows that Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) produces
Raw Substance is thalline secretions, rather than thalline intracellular organic matter, and this Substance molecular weight is relatively big or with
Presented in molecule aggregate, it is impossible to through 0.22um microporous filter membrane；Centrifuged supernatant after heat treatment loses antibacterial work
Property, this shows this Substance non-refractory, easy in inactivation under the high temperature conditions.
Four, the extraction of Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) antipathogenic composition
Ethanol precipitation: be centrifuged at Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36)
Adding dehydrated alcohol in liquid makes the final concentration of ethanol be respectively 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,
Standing 2h, 5000rpm in 4 DEG C and be centrifuged 10min, precipitation be dissolved in sterilized water, gained solution is Paenibacillus polymyxa generation
Thank to (Paenibacillus polymyxa strain DSM 36) protein crude extract administration of product, survey by the method for above-mentioned experiment three
Determine the bacteriostatic activity of different concentration ethanol protein precipitation crude extract.
The bacteriostatic activity of the different determining alcohol extraction of substance of table 2
The protein crude extract administration bacteriostatic activity that table 2 result display concentration of alcohol extracts when being 70% is maximum
Five, Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) field control Caulis Sacchari sinensis phoenix
Pears disease effect experimental
Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) field control Caulis Sacchari sinensis Fructus Ananadis comosi
Sick effect experimental sets 4 process: Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) bacterium is hanged
1000 times of diluents of liquid, effective bacteria concentration is 104Cfu/ml (preparation method sees the embodiment of the present application 1)；Paenibacillus
Polymyxa strain DSM 36 bacteria suspension alcohol extract (its preparation method sees the embodiment of the present application 7)；Carbendazim is wettable
Property powder (being purchased from Sichuan Guoguang Agrochemical Co., Ltd.) 1000 times of diluents；Sterilized water blank.By a length of 20cm
Sugarcane kind pulls neat stacking out after soaking 2h in 3% lime water, the otch of sugarcane kind is placed in approximately the same plane, on otch respectively
After spraying above-mentioned 4 kinds of testing samples, sugarcane kind is planted on arable land, each process 100 strain；Within 1 month, Caulis Sacchari sinensis neat Seedling " Invest, Then Investigate " is sprouted
Bud rate, then takes 40 double sprouts at random from each test treatment region, and with cutter, each sugarcane kind stem is made vertical profile, investigate every
Caulis Sacchari sinensis kind stem pineapple disease incidence, and calculate sickness rate and prevention effect, its result see table:
Table 3 respectively processes the effect of preventing and treating Caulis Sacchari sinensis pineapple disease
The Caulis Sacchari sinensis germination rate of Different treatments, pineapple disease sickness rate, preventive effect result (being shown in Table 3) shows: many viscous class spore
Bacillus (DSM 36) has preferable prevention effect to Caulis Sacchari sinensis pineapple disease, and its preventive effect is 91.3%, carbendazol wettable powder 1000
The preventive effect of times diluent is 52.2%, and the preventive effect of DSM 36 is significantly higher than the anti-of 1000 times of diluents of carbendazol wettable powder
Effect.And the rudiment to sugarcane kind has obvious facilitation.
The invention have the benefit that
1, screening acquisition through microorganism present in nature can the strain of Biological control Caulis Sacchari sinensis pineapple disease, system
Agent and preparation method thereof, solves the problems such as pesticide residues, pathogenic bacteria resistance to drugs and environmental pollution.
2, preparation of the present invention can not only prevent or treat Caulis Sacchari sinensis pineapple disease, moreover it is possible to the rudiment to sugarcane kind is played bright
Aobvious facilitation.
Accompanying drawing explanation
Fig. 1 is the gram of the isolated bacterial strain of the application i.e. Paenibacillus polymyxa strain DSM 36
Dyeing picture；
Fig. 2 is that the antagonistic activity of pineapple disease pathogen is tested by Paenibacillus polymyxa strain DSM 36
Result picture；
Detailed description of the invention
Embodiment 1
A kind of preparation for preventing or treat Caulis Sacchari sinensis pineapple disease, its preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in
Culture medium is cultivated, obtains bacteria suspension；
2) above-mentioned bacteria suspension direct packaging, both.
Embodiment 2
A kind of preparation for preventing or treat Caulis Sacchari sinensis pineapple disease, its preparation method is as follows:
1) take Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36), strain be inoculated in
Culture medium is cultivated, obtains bacteria suspension；
2) above-mentioned bacteria suspension takes supernatant after centrifugal treating, both obtains.
Embodiment 3
A kind of preparation for preventing or treat Caulis Sacchari sinensis pineapple disease, its preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in
In LB fluid medium 30 degrees Celsius, 200rpm concussion is cultivated 24 hours, and obtaining effective bacteria concentration is 103The bacterium of cfu/ml is hanged
Liquid；
2) take above-mentioned bacteria suspension after centrifugal treating, obtain supernatant, supernatant adds ethanol so that determining alcohol
It is 20%, centrifugal after standing, take its precipitation, precipitation is dissolved in sterilized water, be prevention or the system for the treatment of Caulis Sacchari sinensis pineapple disease
Agent.
Prepared preparation, by method described in Summary test three, carries out effect test, and its inhibition zone size is
25.6mm。
Embodiment 4
A kind of preparation for preventing or treat Caulis Sacchari sinensis pineapple disease, its preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in
In LB fluid medium 35 degrees Celsius, 240rpm concussion is cultivated 48 hours, and obtaining effective bacteria concentration is 108The bacterium of cfu/ml is hanged
Liquid；
2) take above-mentioned bacteria suspension after centrifugal treating, obtain supernatant, supernatant adds ethanol so that determining alcohol
It is 90%, centrifugal after standing, take its precipitation, precipitation is dissolved in sterilized water, be prevention or the system for the treatment of Caulis Sacchari sinensis pineapple disease
Agent.
Prepared preparation, by method described in Summary test three, carries out effect test, and its inhibition zone size is
27.4mm。
Embodiment 5
A kind of preparation for preventing or treat Caulis Sacchari sinensis pineapple disease, its preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in
In LB fluid medium 33 degrees Celsius, 220rpm concussion is cultivated 36 hours, and obtaining effective bacteria concentration is 106The bacterium of cfu/ml is hanged
Liquid.
2) take the bacteria suspension obtained in 1 parts by volume step 1, after 10000 are centrifuged 8min, take supernatant and add ethanol so that
Determining alcohol is 60%, and stand at low temperature 2000 was centrifuged 5 after 1 hour, takes its precipitation, precipitation is dissolved in taken bacteria suspension parts by volume 2 times
In the sterilized water of amount, obtain the extract of Paenibacillus polymyxa metabolite, be prevention or treat Caulis Sacchari sinensis pineapple disease
Preparation.
Prepared preparation, by method described in Summary test three, carries out fungistatic effect test, and its inhibition zone is big
Little for 26.9mm.
Embodiment 6
A kind of preparation for preventing or treat Caulis Sacchari sinensis pineapple disease, its preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in
In LB fluid medium 33 degrees Celsius, 220rpm concussion is cultivated 36 hours, and obtaining effective bacteria concentration is 107The bacterium of cfu/ml is hanged
Liquid.
2) take the bacteria suspension obtained in 1 parts by volume step 1, after 14000rpm is centrifuged 12min, takes supernatant and add ethanol,
Making determining alcohol is 80%, stand at low temperature after 3 hours 7000rpm be centrifuged 15min, take its precipitation, precipitation is dissolved in taken bacterium and hangs
In the sterilized water of liquid parts by volume 6 times amount, obtain the extract of Paenibacillus polymyxa metabolite, be prevention or treat sweet
The preparation of sugarcane pineapple disease.
Prepared preparation, by method described in Summary test three, carries out effect test, and its inhibition zone size is
27.3mm。
Embodiment 7
A kind of preparation for preventing or treat Caulis Sacchari sinensis pineapple disease, its preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in
In LB fluid medium 33 degrees Celsius, 220rpm concussion is cultivated 36 hours, and obtaining effective bacteria concentration is 106-107The bacterium of cfu/ml
Suspension.
2) take the bacteria suspension obtained in 1 parts by volume step 1, after 12000rpm is centrifuged 10min, takes supernatant and add ethanol,
Making determining alcohol is 70%, and after standing 2 hours in 4 degrees Celsius, 5000rpm is centrifuged 10min, takes its precipitation, precipitation is dissolved in and being taken
In the sterilized water of bacteria suspension parts by volume 4 times amount, it is prevention or the preparation for the treatment of Caulis Sacchari sinensis pineapple disease.
Prepared preparation, by method described in Summary test three, carries out effect test, and its inhibition zone size is
28.3mm。
Claims (10)
1. one kind for preventing or treat the preparation of Caulis Sacchari sinensis pineapple disease, it is characterised in that said preparation by effective ingredient for glue more
Series bacillus (Paenibacillus polymyxa strain DSM 36) through cultivate after bacteria suspension or this bacterium metabolism
The extract of product is made.
2. preparation as claimed in claim 1, it is characterised in that described extraction and preparation method be: by Paenibacillus polymyxa
Bacteria suspension after cultivating, through its supernatant of centrifuging and taking, both obtained.
3. preparation as claimed in claim 1, it is characterised in that described extraction and preparation method be:
1) taking Paenibacillus polymyxa bacteria suspension 1 parts by volume after cultivating, its effective bacteria concentration is 103-108Cfu/ml,
After 10000-14000rpm is centrifuged 8-12min, take supernatant；
2) in centrifuged supernatant, ethanol is added so that determining alcohol is 20%-90%, centrifugal after standing, take its precipitation, will precipitation
It is dissolved in the sterilized water of taken 2-6 times of parts by volume of bacteria suspension, both.
4. preparation as claimed in claim 1, it is characterised in that described extraction and preparation method be:
1) taking Paenibacillus polymyxa bacteria suspension 1 parts by volume after cultivating, its effective bacteria concentration is 106-107Cfu/ml,
12000rpm takes supernatant after being centrifuged 10min；
2) adding ethanol in centrifuged supernatant so that determining alcohol is 70%, after standing 2 hours in 4 degrees Celsius, 5000rpm is centrifuged
10min, takes its precipitation, precipitation is dissolved in the sterilized water of taken 4 times of parts by volume of bacteria suspension, to obtain final product.
5. the preparation as described in any one of claim 1-4, it is characterised in that described preparation can be used in prevention or treats sweet
Sugarcane pineapple disease.
6. the preparation method being used for preventing or treat the preparation of Caulis Sacchari sinensis pineapple disease, it is characterised in that described preparation method
As follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in cultivation
Base is cultivated, obtains bacteria suspension；
2) above-mentioned bacteria suspension takes supernatant after centrifugal treating, both obtains.
7. preparation method as claimed in claim 6, it is characterised in that described preparation method is as follows:
1) strain taking Paenibacillus polymyxa (Paenibacillus polymyxa strain DSM 36) is inoculated in LB liquid
In body culture medium 30-35 degree Celsius, 200-240rpm concussion is cultivated 24-48 hour, and obtaining effective bacteria concentration is 103-108cfu/
The bacteria suspension of ml；
2) take above-mentioned bacteria suspension after centrifugal treating, obtain supernatant, supernatant adds ethanol so that determining alcohol is
20%-90%, centrifugal after standing, take its precipitation, precipitation is dissolved in sterilized water, be prevention or treat Caulis Sacchari sinensis pineapple disease
Preparation.
8. preparation method as claimed in claim 7, it is characterised in that described step 1 is: take Paenibacillus polymyxa
The strain of (Paenibacillus polymyxa strain DSM 36) is inoculated in LB fluid medium 33 degrees Celsius,
220rpm concussion is cultivated 36 hours, and obtaining effective bacteria concentration is 106-107The bacteria suspension of cfu/ml.
9. preparation method as claimed in claim 7, it is characterised in that described step 2 is: takes in 1 parts by volume step 1 and obtains
Bacteria suspension, after 10000-14000rpm is centrifuged 8-12min, takes supernatant and adds ethanol so that determining alcohol is 60-80%, low temperature
After standing 1-3 hour, 2000-7000rpm is centrifuged 5-15min, takes its precipitation, precipitation is dissolved in taken bacteria suspension parts by volume 2-6 times
In the sterilized water of amount, obtain the extract of Paenibacillus polymyxa metabolite, be prevention or treat Caulis Sacchari sinensis pineapple disease
Preparation.
10. preparation method as claimed in claim 7, it is characterised in that described step 2 is: takes in 1 parts by volume step 1 and obtains
Bacteria suspension, after 12000rpm is centrifuged 10min, take supernatant add ethanol so that determining alcohol is 70%, in 4 degrees Celsius stand 2
After hour, 5000rpm is centrifuged 10min, takes its precipitation, precipitation is dissolved in the sterilized water of taken bacteria suspension parts by volume 4 times amount, i.e.
For prevention or the preparation for the treatment of Caulis Sacchari sinensis pineapple disease.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610424369.4A CN106047965A (en) | 2016-06-15 | 2016-06-15 | Preparation for preventing or treating eratocystis paradoxa and preparation method of preparation |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610424369.4A CN106047965A (en) | 2016-06-15 | 2016-06-15 | Preparation for preventing or treating eratocystis paradoxa and preparation method of preparation |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106047965A true CN106047965A (en) | 2016-10-26 |
Family
ID=57168020
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610424369.4A Pending CN106047965A (en) | 2016-06-15 | 2016-06-15 | Preparation for preventing or treating eratocystis paradoxa and preparation method of preparation |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106047965A (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101712941A (en) * | 2009-11-30 | 2010-05-26 | 浙江大学 | Paenibacillus polymyxa and application thereof |
| US20140030228A1 (en) * | 2012-07-30 | 2014-01-30 | Mineral Biosciences Llc | Methods and Compositions of Biocontrol of Plant Pathogens |
-
2016
- 2016-06-15 CN CN201610424369.4A patent/CN106047965A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101712941A (en) * | 2009-11-30 | 2010-05-26 | 浙江大学 | Paenibacillus polymyxa and application thereof |
| US20140030228A1 (en) * | 2012-07-30 | 2014-01-30 | Mineral Biosciences Llc | Methods and Compositions of Biocontrol of Plant Pathogens |
Non-Patent Citations (2)
| Title |
|---|
| M. A. RAHMAN等: "Screening of Trichoderma Isolates as a Biological Control Agent against Ceratocystis paradoxa Causing Pineapple Disease of Sugarcane", 《MYCOBIOLOGY》 * |
| 霍秀娟等: "广西甘蔗病害种类调查与鉴定", 《安徽农业科学》 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105886428B (en) | One plant of Streptomycesalbidoflhaving and its application in microbial manure | |
| CN100334201C (en) | Bacillus subtilis and its uses | |
| CN111254086B (en) | Bacillus belgii and application thereof in biocontrol | |
| CN103451135B (en) | Bacillus subtilis M3 and application thereof | |
| CN110079483A (en) | A kind of bacillus subtilis and its application | |
| CN103074271B (en) | Bacillus subtilis YB-05, microbial preparation thereof, and application of Bacillus subtilis YB-05 or microbial preparation | |
| CN102433282A (en) | Bacillus subtilis NB12, as well as culture method and application thereof | |
| CN109136157A (en) | One plant of Bei Laisi bacillus for preventing and treating rice blast and its application | |
| CN104611254B (en) | Aerobacillus polymyxa Donker KN 03 and its cultural method and purposes | |
| CN106942301B (en) | A kind of botanical pesticide preparation of prevention and control rice blast and preparation method thereof | |
| CN103509740A (en) | Pseudomonas chlororaphis for preventing and treating crop fusarium disease and applications thereof | |
| CN102533617A (en) | Bacillus subtilis strain and application thereof | |
| KR20120030630A (en) | Bacillus amyloliquefaciens jbc36 and composition comprising the same for control of plant pathogens | |
| CN105368720A (en) | Cotton fungal endophyte CEF-082 and application of cotton fungal endophyte CEF-082 in prevention and treatment of cotton verticillium wilt | |
| CN101518264B (en) | Microbial preparation for preventing and controlling plant silborne fungal diseases, and application thereof | |
| CN103305438A (en) | Biocontrol strain Jdm2 for preventing and treating tomato root-knot nematode diseases and antibiological inoculant thereof | |
| CN102925387A (en) | Bacillus simplex for inducing soybean to generate soybean cyst nematode resistance and application | |
| CN106010983B (en) | Cotton endogenetic fungus CEF-559 and its application in cotton verticillium wilt prevention and treatment | |
| CN102443559B (en) | Baclillus subtilis used for controlling cotton verticillium wilt and application thereof | |
| CN102732430A (en) | Aspergillus niger strain and application thereof | |
| CN106676047A (en) | Bacillus amyloliquefaciens HM-6, and preparation method and application of microbial inoculum thereof | |
| WO2002034884A1 (en) | Streptomyces kasugaensis inhibiting the fungal pathogens of plant | |
| CN1327774C (en) | Tobacco rust resisting microbial prepn and its prepn process and application | |
| CN115141785A (en) | Bacillus subtilis and application thereof in cabbage planting | |
| CN109207392A (en) | One plant of bacillus megaterium and its application in prevention and treatment American ginseng root maize ear rot |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161026 |