CN108192829A - A kind of gamboge streptomyces verticillus and its biological control agent and application - Google Patents
A kind of gamboge streptomyces verticillus and its biological control agent and application Download PDFInfo
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- CN108192829A CN108192829A CN201810032717.2A CN201810032717A CN108192829A CN 108192829 A CN108192829 A CN 108192829A CN 201810032717 A CN201810032717 A CN 201810032717A CN 108192829 A CN108192829 A CN 108192829A
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- 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
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- 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
- C12R2001/465—Streptomyces
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- 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
- A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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
- C12N1/14—Fungi; Culture media therefor
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- 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
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
Abstract
The invention discloses a kind of gamboge streptomyces verticillus and its biological control agent and applications, belong to biological control plant disease technical field, gamboge streptomyces verticillus bacterial strain HY61 is located away from Yantai marine site bottom sediment, China Committee for Culture Collection of Microorganisms's common micro-organisms center is preserved in, deposit number is CGMCC No.15060.It is strong that the gamboge streptomyces verticillus bacterial strain HY61 of the present invention has many advantages, such as production, and spore speed is fast, action spectrum extensively prevents plurality of plant diseases, genetic stability, and environment compatibility is good, the diseases prevention lasting period is long, therefore has a good application prospect.
Description
Technical field
The present invention relates to biological control plant disease technical field, more particularly to a kind of gamboge streptomyces verticillus and its biology
Prevent preparation and application.
Background technology
Plant disease is the antagonistic symbiosis of host plant and pathogen, occurs and prevalence is host plant and pathogen
The result of interaction.The disease of crops and forest occurs, and national economy and people's lives is often made to sustain losses severely.In recent years
Come, as the public is to the common concern of chemical pesticide safety in food and environment and the taboo of some dangerous chemical pesticides
With people is promoted to seek other safely and effectively plant disease control new ways, utilize beneficial microbe controlling plant diseases day
Become to being taken seriously.It is that the external source Antagonistic Fungi beneficial to plant is introduced into the plant natural ecosystem being broken or is passed through
Adjusting ambient promotes natural beneficial microbe group to increase and shows biological and ecological methods to prevent plant disease, pests, and erosion activity, is reached by the interaction between microorganism
To the purpose of diseases prevention.
Actinomyces are widely present in nature, most medium temperatures, saprophytic, a small number of high temperature, thermophilic salt, acidophilus, basophilic, anaerobism,
Parasitism is mainly grown in land and fresh water, and water content is relatively low, is also deposited in air in preferable neutral or meta-alkalescence soil of ventilating
.Report about the biocontrol actinomycetes of the tool control of plant disease of separation screening from soil has very much, Yan Jianfang etc. from
Isolated more plants of actinomyces in the soil in Xinjiang and Liaoning, screening obtain the actinomyces for having antagonism to Fusarium oxysporum
Bacterial strain.Sun Qiang etc. screens the unwrapping wire for being separated to one plant of streptomyces rimosus cracking subspecies from the soil of the Northeast's vegetable protecting field
Bacteria strain, result of the test show that its zymotic fluid has antifungal activity.
Marine environment is special, has the characteristics that high salinity, high pressure, low temperature, poor nutritional, survives in this particular surroundings
Marine actinomycete, there is the unique metabolic approach different from terrestrial actinomyces.Microorganism novel species and tool are found from ocean
The physiological activator for having specific function becomes research hotspot both domestic and external, and marine microorganism has become agricultural disease resistance activity
The new resources of substance.Recent study finds that more than 50% newfound research of bioactive substances from marine microorganisms is produced by marine actinomycete
Raw, the secondary metabolite of many actinomyces has the purposes in terms of medical and plant protection, has been widely used as antibacterium, has resisted
Plant pathogenic fungi and antitumor drug.
Invention content
The present invention provides a kind of gamboge streptomyces verticillus and its biological control agent and application, gamboge of the invention wheel silks
Streptomyces strain has the characteristics that biocontrol effect is good, the wide anti-various plants pathogen of action spectrum.
Technical scheme is as follows：
Gamboge streptomyces verticillus bacterial strain HY61 provided by the invention, is isolated from Yantai marine site bottom sediment, in being preserved in
State's General Microbiological Culture preservation administrative center, preservation date are on December 13rd, 2017, and Classification And Nomenclature takes turns silk strepto- for gamboge
Bacterium (Streptomyces luteoverticillatus), deposit number is：CGMCC No.15060.
Further, the 16S rRNA gene orders such as SEQ.NO.1 of the bacterial strain.
The present invention provides gamboge streptomyces verticillus (Streptomyces luteoverticillatus) bacterial strain HY61 to exist
Application in biological control plant disease.
The present invention provides a kind of biological control agent, including the gamboge streptomyces verticillus (Streptomyces
Luteoverticillatus) bacterial strain HY61 and its metabolite.
The present invention provides a kind of method for preparing biological control agent, includes the following steps：
(1) bacterial strain activates：The streak inoculation of picking strain is to solid slope culture medium, in 25~32 DEG C of cultures of constant incubator
3~7d treats that inclined-plane grows a large amount of spores for liquid seeds of transferring；
(2) prepared by liquid seeds：Picking activated strains are inoculated in the container equipped with seed culture medium, gauze sealing, in
25-32 DEG C, 150~200r/min shaking table 24~48h of shaken cultivation grow a large amount of mycelium pellets or mycelium, obtain liquid seeds；
(3) liquid fermentation：The liquid seeds that 1~15% inoculum concentration is prepared to fermentation medium access step 2 by volume,
200~700r/min of rotating speed, dissolved oxygen control volume ratio 10~40%, pH6.0~7.5,25~32 DEG C of temperature, ventilation culture 48~
72h, zymotic fluid can be used as biological control agent.
Further, the ingredient and final concentration of the solid slope culture medium：Yeast extract 4g/L, fructus hordei germinatus leaching powder 10g/L,
Glucose 4g/L, 18g/L, pH7.0,121 DEG C of high pressure steam sterilization 20min of agar；
Further, the ingredient and final concentration of the liquid seed culture medium：Yeast extract 4g/L, fructus hordei germinatus leaching powder 10g/L,
4g/L, pH7.0,121 DEG C of high pressure steam sterilization 20min of glucose；
Further, the ingredient and final concentration of the fermentation medium：Starch 15g/L, glycerine 5g/L, peptone 11g/L,
Yeast extract 4.4g/L, potassium nitrate 0.4g/L, magnesium sulfate 4g/L, KH2PO40.2g/L, KCl 0.2g/L, CaCl210mg/L,
VB10.5mg/L, riboflavin 0.5mg/L, VH0.5mg/L, niacin 1mg/L, FeSO40.1mg/L, VB120.5mg/L, pH
7.0,121 DEG C of high pressure steam sterilization 20min.
The present invention provides application of the biological control agent in controlling plant diseases drug.The phytopathy
Evil includes colletotrichum gloeosporioides Penz (Colletotrichum gloeosporioides), the pathogen of Botrytis cinerea (Botrytis
Cinerea), Fusarium oxysporum (Fusarium oxysporum), gibberella saubinetii (Fusarium graminearum
Schw.), maize curvularia (Curvularia lunata) etc..
The advantageous effect of the present invention compared with prior art：The gamboge streptomyces verticillus bacterial strain HY61 of the present invention has production spore
It is strong that speed is fast, action spectrum extensively prevents plurality of plant diseases, genetic stability, excellent with good, the diseases prevention lasting period length of environment compatibility etc.
Point, therefore have a good application prospect.
Description of the drawings
Fig. 1 gamboge streptomyces verticillus bacterial strain HY61 colonial morphologies
Phylogenetic trees of the Fig. 2 based on 16S rRNA sequences
Gamboge streptomyces verticillus bacterial strain HY61, is isolated from Yantai marine site bottom sediment, is preserved in China General Microbiological
Culture presevation administrative center, preservation date are on December 13rd, 2017, and Classification And Nomenclature is gamboge streptomyces verticillus
(Streptomyces luteoverticillatus), deposit number is：CGMCC No.15060.
Specific embodiment
To illustrate the understanding to this invention feature, the present invention is done into one with reference to some non-limiting embodiments
Step illustrates, but does not therefore limit the present invention among the embodiment described range.
Embodiment 1：Bacterial strain HY61 Isolation and screening of bacterial strain
1) it detaches：The bacterial strain HY61 of the present invention is obtained using method of dilution butteron on plate separation, and specific separation method is：It weighs
10g bottom sediment samples are dissolved in 90mL sterile waters, and oscillation, which shakes up, makes sample be sufficiently mixed with water, is coated on after gradient dilution
In Gause I culture medium flat plate.Tablet is placed in 28 DEG C of insulating box and cultivates 5d, by the bacterium colony of the different shape on culture medium
It crosses in blank Gause I culture medium flat plate, purifies the bacterial strain of acquisition, and number preserves respectively.Wherein Gause I
Nutrient media components are：Soluble starch 20g/L, KNO31g/L, K2HPO40.5g/L, MgSO40.5g/L, NaCl 0.5g/L,
FeSO40.01g/L, 20g/L, pH7.4,121 DEG C of high pressure steam sterilization 20min of agar.
2) it screens：Using opposite culture method, maize curvularia is inoculated into PDA culture medium with the card punch of diameter 5mm
Midpoint is inoculated with 10 μ L antagonism bacterium solutions from symmetrical both sides at the 2cm of center in fungus block, is inverted, in 28 DEG C of constant temperature incubation 5d, not connect
The culture dish of kind Antagonistic Fungi compares.Wait compare ware mycelia cover with culture dish when checkout facility result.Wherein PDA culture medium component
For：Potato 200g/L, glucose 20g/L, agar 20g/L, pH are naturally, 121 DEG C of high pressure steam sterilization 20min.
One plant of bacterial strain HY61 that can efficiently prevent various plants pathogen disease, experiment card are obtained by a large amount of screening operations
The bright biological control agent prepared by the bacterium is a kind of biological control agent of great application prospect.
Embodiment 2：Bacterial strain HY61 bacterial strains are identified
1) strain properties
With colonial morphology circle, bacterium colony surface shrinkage (see Fig. 1), powder after Gause I medium culture bacterial strain HY61,5d
Shape, it is near greyish white, there is mud fishy smell；The raw bacterium white of gas, substrate mycelium light brown is yellow, secretes soluble brown pigment.In the height of inserted sheet observation
On family name's No.1 culture medium flat plate, aerial hyphae forms longer axial filament, and interval grows branch, and two level is verticillate, and fibrillae of spores is ripe
After fragment into spore, spore is in elliposoidal, the bending of substrate mycelium branch.
Bacterial strain HY61 being capable of decomposition glucose, fructose, glycerine and starch, it is impossible to be gathered using sucrose, lactose, cellulose and shell
Sugar.The result of the tests such as milk solidification, pigment generation, gelatin liquefaction are that the result of the tests such as positive, methyl red and casein hydrolysis are
It is negative.Well-grown in 3%NaCl culture mediums, result of the test refer to table 1.
1 bacterial strain HY61 physiological and biochemical properties of table
2) bacterial strain is identified
Extracted through DNA, 16S rRNA gene orders length that PCR amplification and sequencing obtain is 1430bp, on GenBank
BLAST comparisons are carried out, obtain the bacterial strain and Streptomyces luteoverticillatus (NR118282) base similitude
Up to 99%, HY61 bacterial strains are accredited as gamboge streptomyces verticillus (Streptomyces by combining form and physiological and biochemical index
Luteoverticillatus) (see Fig. 2).16S rRNA sequence informations are as follows：
caacgggcagtcaccttcgacggctcctccacaagggttgggccaccggcttcgggtgttaccgactttcgtgacgt
gacgggcggtgtgtacaaggcccgggaacgtattcaccgcagcaatgctgatctgcgattactagcaactccaactt
catggggtcgagttgcagaccccaatccgaactgagaccggctttttgagattcgctccacctcacggcatcgcagc
tcattgtaccggccattgtagcacgtgtgcagcccaagacataaggggcatgatgacttgacgtcgtccccaccttc
ctccgagttgaccccggcagtctcctgtgagtccccatcaccccgaagggcatgctggcaacacaggacaagggttg
cgctcgttgcgggacttaacccaacatctcacgacacgagctgacgacagccatgcaccacctgtacaccgaccaca
agggggcgaccatctctggccgtttccggtgtatgtcaagccttggtaaggttcttcgcgttgcgtcgaattaagcc
acatgctccgctgcttgtgcgggcccccgtcaattcctttgagttttagccttgcggccgtactccccaggcgggga
acttaatgcgttagctgcggcacggacgacgtggaatgtcgcccacacctagttcccaacgtttacggcgtggacta
ccagggtatctaatcctgttcgctccccacgctttcgctcctcagcgtcagtatcggcccagagatccgccttcgcc
accggtgttcctcctgatatctgcgcatttcaccgctacaccaggaattccgatctcccctaccgaactctagcctg
cccgtatcgaatgcagacccggggttaagccccgggctttcacatccgacgcgacaagccgcctacgagctctttac
gcccaataattccggacaacgcttgcgccctacgtattaccgcggctgctggcacgtagttagccggcgcttcttct
gcaggtaccgtcactttcgcttcttccctgctgaaagaggtttacaacccgaaggccgtcatccctcacgcggcgtc
gctgcatcaggctttcgcccattgtgcaatattccccactgctgcctcccgtaggagtctgggccgtgtctcagtcc
cagtgtggccggtcgccctctcaggccggctacccgtcgtcgccttggtaggccatcaccccaccaacaagctgata
ggccgcgggctcatcctgcaccgccggagctttccaccaccaaccatgcggtcagtagtcatatccggtattagacc
ccgtttccagggcttgtcccagagtgcagggcagattgcccacgtgttactcacccgttcgccactaatccaccccg
aagggcttcatcgttcgactgcatggtagccccccgcacggggc
3) genetic stability
Using method of scoring continuous passage 50 times in solid slope culture medium by gamboge streptomyces verticillus HY61, thalline is measured
The ability of form, the speed of growth and anti-maize curvularia, with primary bacterial strain no significant difference, genetic stability is good.
4) culture presevation
Gamboge streptomyces verticillus (Streptomyces luteoverticillatus) HY61 described in this invention,
China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number are preserved on December 13rd, 2017：
CGMCC No.15060, depositary institution address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences microbe research
Institute.
3 gamboge streptomyces verticillus HY61 fermentation process of embodiment
(1) bacterial strain activates：The streak inoculation of picking strain is to solid slope culture medium：Yeast extract 4g/L, fructus hordei germinatus leaching powder
10g/L, glucose 4g/L, 18g/L, pH7.0,121 DEG C of high pressure steam sterilization 20min of agar, in 28 DEG C of cultures of constant incubator
7d treats that inclined-plane grows a large amount of spores for liquid seeds of transferring；
(2) prepared by liquid seeds：Picking activated strains are inoculated in equipped with seed culture medium (yeast extract 4g/L, malt leaching
Powder 10g/L, glucose 4g/L, pH7.0,121 DEG C high pressure steam sterilization 20min) in, gauze sealing, in 30 DEG C, 200r/min shakes
Bed shaken cultivation 48h, grows a large amount of mycelium pellets or mycelium, obtains liquid seeds；
(3) liquid fermentation：1~15% inoculum concentration is to fermentation medium (starch 15g/L, glycerine 5g/L, albumen by volume
Peptone 11g/L, yeast extract 4.4g/L, potassium nitrate 0.4g/L, magnesium sulfate 4g/L, KH2PO40.2g/L, KCl 0.2g/L, CaCl2
10mg/L, VB10.5mg/L, riboflavin 0.5mg/L, VH0.5mg/L, niacin 1mg/L, FeSO40.1mg/L, VB12 0.5mg/
7.0,121 DEG C of high pressure steam sterilization 20min of L, pH) access liquid seeds prepared by step 2, rotating speed 500r/min, dissolved oxygen control
20%, pH7.0,30 DEG C of temperature, ventilation culture 72h, you can as biological control agent.
The 4 disease-resistant experiment of gamboge streptomyces verticillus HY61 zymotic fluids of embodiment
The inspection of the anti-phytopathogen ability of gamboge streptomyces verticillus HY61 zymotic fluids is carried out by cylinder-plate method, as a result such as table
Shown in 2.Gamboge streptomyces verticillus HY61 zymotic fluids are to colletotrichum gloeosporioides Penz (Colletotrichum gloeosporioides), ash
Botrytis (Botrytis cinerea), Fusarium oxysporum (Fusarium oxysporum), gibberella saubinetii (Fusarium
Graminearum Schw.), the growths of the phytopathogens such as maize curvularia (Curvularia lunata) is respectively provided with significantly
Antagonism.Wherein the most notable to the inhibition of maize curvularia, the average inhibition diameter of 10 μ L zymotic fluids reaches
4cm；Good inhibiting effect has also been shown to other phytopathogens, has averagely inhibited diameter that in more than 2.0cm, all there is exploitation
The potentiality utilized.
2 bacterial strain HY61 zymotic fluids of table inhibit the antibacterial circle diameter and area of phytopathogen
5 gamboge streptomyces verticillus HY61 zymotic fluids of embodiment verify Curvularia lunata field efficacy
With No. 5 syringe needles, blade pierces 6 needles in the middle part of corn, and Curvularia lunata is then inoculated at needle thorn,
50 plants are often handled, 3 repetitions spray 500 times of dilutions of gamboge streptomyces verticillus HY61 zymotic fluids, 50% carbendazim 500 after 2d
Times liquid and not dispenser clear water compare totally 3 processing, and 7 days investigation disease indexs, calculate control effect after dispenser.Result of the test
(table 3) shows after dispenser 7 days, and 500 times of dilution control effects of gamboge streptomyces verticillus HY61 zymotic fluids are 73.2%, significantly
Higher than the effect of 50% 500 times of liquid of carbendazim.
3 gamboge streptomyces verticillus HY61 zymotic fluids of table are to the control effect of Curvularia lunata
Sequence table
<110>Ludong University
<120>A kind of gamboge streptomyces verticillus and its biological control agent and application
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1430
<212> DNA
<213>Gamboge streptomyces verticillus (Streptomyces luteoverticillatus)
<400> 1
caacgggcag tcaccttcga cggctcctcc acaagggttg ggccaccggc ttcgggtgtt 60
accgactttc gtgacgtgac gggcggtgtg tacaaggccc gggaacgtat tcaccgcagc 120
aatgctgatc tgcgattact agcaactcca acttcatggg gtcgagttgc agaccccaat 180
ccgaactgag accggctttt tgagattcgc tccacctcac ggcatcgcag ctcattgtac 240
cggccattgt agcacgtgtg cagcccaaga cataaggggc atgatgactt gacgtcgtcc 300
ccaccttcct ccgagttgac cccggcagtc tcctgtgagt ccccatcacc ccgaagggca 360
tgctggcaac acaggacaag ggttgcgctc gttgcgggac ttaacccaac atctcacgac 420
acgagctgac gacagccatg caccacctgt acaccgacca caagggggcg accatctctg 480
gccgtttccg gtgtatgtca agccttggta aggttcttcg cgttgcgtcg aattaagcca 540
catgctccgc tgcttgtgcg ggcccccgtc aattcctttg agttttagcc ttgcggccgt 600
actccccagg cggggaactt aatgcgttag ctgcggcacg gacgacgtgg aatgtcgccc 660
acacctagtt cccaacgttt acggcgtgga ctaccagggt atctaatcct gttcgctccc 720
cacgctttcg ctcctcagcg tcagtatcgg cccagagatc cgccttcgcc accggtgttc 780
ctcctgatat ctgcgcattt caccgctaca ccaggaattc cgatctcccc taccgaactc 840
tagcctgccc gtatcgaatg cagacccggg gttaagcccc gggctttcac atccgacgcg 900
acaagccgcc tacgagctct ttacgcccaa taattccgga caacgcttgc gccctacgta 960
ttaccgcggc tgctggcacg tagttagccg gcgcttcttc tgcaggtacc gtcactttcg 1020
cttcttccct gctgaaagag gtttacaacc cgaaggccgt catccctcac gcggcgtcgc 1080
tgcatcaggc tttcgcccat tgtgcaatat tccccactgc tgcctcccgt aggagtctgg 1140
gccgtgtctc agtcccagtg tggccggtcg ccctctcagg ccggctaccc gtcgtcgcct 1200
tggtaggcca tcaccccacc aacaagctga taggccgcgg gctcatcctg caccgccgga 1260
gctttccacc accaaccatg cggtcagtag tcatatccgg tattagaccc cgtttccagg 1320
gcttgtccca gagtgcaggg cagattgccc acgtgttact cacccgttcg ccactaatcc 1380
accccgaagg gcttcatcgt tcgactgcat ggtagccccc cgcacggggc 1430
Claims (9)
1. a kind of gamboge streptomyces verticillus bacterial strain HY61, it is characterised in that the bacterial strain HY61 is preserved in China General Microbiological bacterium
Kind preservation administrative center, preservation date are on December 13rd, 2017, and Classification And Nomenclature is gamboge streptomyces verticillus, and deposit number is：
CGMCC No.15060。
A kind of 2. gamboge streptomyces verticillus bacterial strain HY61 according to claim 1, it is characterised in that the 16S of bacterial strain HY61
RRNA gene orders such as SEQ.NO.1.
3. a kind of biological control agent, including gamboge streptomyces verticillus bacterial strain HY61 described in claim 1 and its metabolite.
4. the preparation method of a kind of biological control agent described in claim 3, it is characterised in that it includes the following steps：
(1) bacterial strain activates：Picking gamboge streptomyces verticillus bacterial strain HY61 streak inoculations described in claim 1 to solid slope is trained
Base is supported, in 25~32 DEG C of 3~7d of culture of constant incubator, treats that inclined-plane grows a large amount of spores for liquid seeds of transferring；
(2) prepared by liquid seeds：The gamboge streptomyces verticillus bacterial strain HY61 of picking step (1) activation is inoculated in equipped with seed culture
In the container of base, gauze sealing, in 25-32 DEG C, 150~200r/min shaking table 24~48h of shaken cultivation grow a large amount of mycelium pellets
Or mycelium, obtain liquid seeds；
(3) liquid fermentation：The liquid seeds that 1~15% inoculum concentration is prepared to fermentation medium access step (2) by volume turn
200~700r/min of speed, dissolved oxygen control volume ratio 10~40%, pH6.0~7.5,25~32 DEG C of temperature, ventilation culture 48~
72h, zymotic fluid are biological control agent.
A kind of 5. preparation method of biological control agent described in claim 4, it is characterised in that the solid slope culture
The ingredient and final concentration of base：Yeast extract 4g/L, fructus hordei germinatus leaching powder 10g/L, glucose 4g/L, agar 18g/L, pH7.0,121 DEG C
High pressure steam sterilization 20min.
A kind of 6. preparation method of biological control agent described in claim 4, it is characterised in that the liquid seeds culture
The ingredient and final concentration of base：Yeast extract 4g/L, fructus hordei germinatus leaching powder 10g/L, 4g/L, pH7.0,121 DEG C of high steams of glucose go out
Bacterium 20min.
7. the preparation method of a kind of biological control agent described in claim 4, it is characterised in that the fermentation medium
Ingredient and final concentration：Starch 15g/L, glycerine 5g/L, peptone 11g/L, yeast extract 4.4g/L, potassium nitrate 0.4g/L, sulfuric acid
Magnesium 4g/L, KH2PO40.2g/L, KCl 0.2g/L, CaCl210mg/L, VB10.5mg/L, riboflavin 0.5mg/L, VH
0.5mg/L, niacin 1mg/L, FeSO40.1mg/L, VB127.0,121 DEG C of high pressure steam sterilization 20min of 0.5mg/L, pH.
8. application of the biological control agent in controlling plant diseases drug described in claim 3.
9. plant disease according to any one of claims 8 includes colletotrichum gloeosporioides Penz, the pathogen of Botrytis cinerea, Fusarium oxysporum, gibberella saubinetii
And maize curvularia.
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| CN201810032717.2A CN108192829B (en) | 2018-01-13 | 2018-01-13 | Streptomyces luteorusand biological control preparation and application thereof |
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| CN201810032717.2A CN108192829B (en) | 2018-01-13 | 2018-01-13 | Streptomyces luteorusand biological control preparation and application thereof |
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| CN108192829B CN108192829B (en) | 2021-03-02 |
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| CN201810032717.2A Active CN108192829B (en) | 2018-01-13 | 2018-01-13 | Streptomyces luteorusand biological control preparation and application thereof |
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Cited By (5)
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| CN109536543A (en) * | 2018-12-25 | 2019-03-29 | 鲁东大学 | A kind of method that microbe fermentation method prepares carbazomycin B |
| CN109576200A (en) * | 2019-01-10 | 2019-04-05 | 鲁东大学 | A kind of recombinant bacterium producing glutamate racemase and its construction method and application |
| CN112899181A (en) * | 2020-11-26 | 2021-06-04 | 湖南农业大学 | Actinomycetes with inhibition effect on peanut soil-borne pathogenic fungi and screening method thereof |
| CN113621526A (en) * | 2021-09-07 | 2021-11-09 | 鲁东大学 | Marine fungus Aspergillus versicolor M-7-SW9, mixed source terpenoid and extraction method and application thereof |
| US11457634B2 (en) | 2019-08-01 | 2022-10-04 | Zhejiang Normal University | S. roseoverticillatus Sr-63 and its application |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109536543A (en) * | 2018-12-25 | 2019-03-29 | 鲁东大学 | A kind of method that microbe fermentation method prepares carbazomycin B |
| CN109576200A (en) * | 2019-01-10 | 2019-04-05 | 鲁东大学 | A kind of recombinant bacterium producing glutamate racemase and its construction method and application |
| US11457634B2 (en) | 2019-08-01 | 2022-10-04 | Zhejiang Normal University | S. roseoverticillatus Sr-63 and its application |
| CN112899181A (en) * | 2020-11-26 | 2021-06-04 | 湖南农业大学 | Actinomycetes with inhibition effect on peanut soil-borne pathogenic fungi and screening method thereof |
| CN112899181B (en) * | 2020-11-26 | 2022-05-06 | 湖南农业大学 | Actinomycetes with inhibition effect on peanut soil-borne pathogenic fungi and screening method thereof |
| CN113621526A (en) * | 2021-09-07 | 2021-11-09 | 鲁东大学 | Marine fungus Aspergillus versicolor M-7-SW9, mixed source terpenoid and extraction method and application thereof |
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| CN108192829B (en) | 2021-03-02 |
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