CN106399152A - Streptomyces having inhibitory effect on phytophthorasojae root rot fungi, biocontrol preparation and preparation method of biocontrol preparation - Google Patents
Streptomyces having inhibitory effect on phytophthorasojae root rot fungi, biocontrol preparation and preparation method of biocontrol preparation Download PDFInfo
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Abstract
The invention discloses streptomyces having an inhibitory effect on phytophthorasojae root rot fungi, a biocontrol preparation and a preparation method of the biocontrol preparation, and belongs to the field of agricultural biotechnology. The strain (streptomyces) is streptomyces dorenlacusensis M1-4. The biocontrol preparation is obtained by inoculating a fermentation medium, which mainly takes soybean cake meal and wheat bran as main raw materials, with the streptomyces dorenlacusensis M1-4 which is subject to enlarged cultivation and by conducting fermentation culture, wherein the total number of spores of the biocontrol preparation is greater than or equal to 10<10> spores/g. From the key problem of preventing and controlling soybean diseases by virtue of bio-pesticide antagonistic microbe, the streptomyces dorenlacusensis M1-4 biocontrol preparation, which is capable of significantly relieving the occurrence of phytophthorasojae root rot, is researched and prepared, and the field preventing and controlling rate of the biocontrol preparation reaches 60% or above. With the application of the biocontrol preparation, the growth of soybean rhizosphere pathogenic microorganisms can be inhibited. Meanwhile, the biocontrol preparation can greatly reduce cost and can achieve a relatively good preventing effect.
Description
Technical field
The invention belongs to agricultural biological technical field is and in particular to a kind of streptomycete, biological prevention and control agent and preparation method thereof.
Background technology
Semen sojae atricolor (Glycine max) belongs to important industrial crops, containing abundant phytoprotein, for doing various bean systems
Product, squeeze Oleum Glycines, make soy sauce and extract protein.Bean dregs or wear into the Semen sojae atricolor of coarse powder and be also commonly used for poultry and livestock feed.Semen sojae atricolor gives birth to
Encroached on by multiple diseases in product.Soybean blight is soybean phytophthora root rot again, by soybean phytophthora (Phytophthorasojae)
Infect the oomycetes disease leading to, be that a kind of spread speed is fast, hazard area is big, be difficult the multi cycle soil-borne disease prevented and treated.It is big
The dangerous disease that bean produces, is acknowledged as the second largest serious soybean diseases after soybean cyst nematode evil.By
Seedling stage dead plant rate that soybean phytophthora root rot causes is light by then 10%, heavy then 80-90%, and land for growing field crops occurs this disease or even causes partly
Block has no harvest.Carry out chemical prevention as its Main Means currently with metalaxyl, but prevention effect is undesirable, Semen sojae atricolor epidemic disease simultaneously
The mould resistance problems to metalaxyl are also increasingly serious.It is the weight of integrated control soil-borne disease using soil microorganism controlling disease
Want one of measure.The Antagonistic Actinomycetes streptomyces hygroscopicuses having for prevention and control continuous cropping Strawberry Root Rot of report
(Streptomyces hygroscopicus) and its biological prevention and control agent (CN 104403976A), for cucumber fusarium axysporum preventing and treating
The fermentation of streptomyces albus (Streptomyces albus) and S. tenebrarius (Streptomyces tenebrarius) is produced
Thing (CN 104342389 A), leads to for preventing and treating phytophthora infestans, Fructus Cucumidis sativi phytophthora, Phytophthora capsici or tobacco black shank bacterium
The tunning (CN 103361294 A) of the wet streptomycete (Streptomyces humidus) of disease.
Biological prevention have environmental friendliness, safely, be not likely to produce Drug resistance and multiple advantages such as efficiently, be to realize agriculture
One of continuable effective ways of industry, preventing and treating root rot using biological preparation is current study hotspot.Current comparison
Good root rot biocontrol agent, mainly biological preparation angstrom thumb secretes (a kind of actinomycetes granule), prevents and treats Fusarium spp. and purple
Penicillium；Paecilomyces lilacinus microbial inoculum, the fungal disease such as preventing and treating Fusarium spp..Prevent and treat the report of oomycetes disease currently with drappus microbial inoculum
Road is simultaneously few, and the drappus microbial inoculum being especially effectively directed to soybean phytophthora is also seldom reported.
Content of the invention
In order to solve the technical problem proposing in background technology, present invention offer is a kind of to be had to soybean phytophthora root rot pathogenic bacteria
There are streptomycete, biological prevention and control agent of inhibitory action and preparation method thereof.
This streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria is Duolun lake streptomycete (Streptomyces
Dorenlacusensis) M1-4, this bacterial strain is deposited in China General Microbiological culture presevation administrative center, and preservation address is north
Jing Shi Chaoyang District North Star West Road 1 institute 3, preserving number is CGMCC NO.11789, and preservation date is on December 4th, 2015.
The biological prevention and control agent of the described streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria it is characterised in that：
This biological prevention and control agent is to expand Duolun lake streptomycete (Streptomyces dorenlacusensis) M1-4 described in claim 1
Big cultivate after be inoculated in the fermentation medium with soybean cake powder and Testa Tritici as primary raw material and carry out fermentation culture to obtain spore total
Number >=1010The biological prevention and control agent of individual/g.
The preparation method bag of the described biological prevention and control agent of streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria
Include following steps：(1) amplification culture of strain：Take Duolun lake streptomycete (Streptomyces dorenlacusensis) M1-4
The conidium of bacterial strain, is inoculated in liquid seed culture medium, and shaking table culture obtains streptomycete liquid seeds, the bar of shaking table culture
Part is 28～30 DEG C of temperature, rotating speed 180r/min, incubation time 48h；The formula of described liquid seed culture medium is：Yeast carries
Take thing 4g/L, glucose 4g/L, Fructus Hordei Germinatus extract 10g/L, CaCO32g/L, pH value 7.2～7.4；(2) system of biological prevention and control agent
Standby：By soybean cake powder and Testa Tritici in mass ratio 1：Moisture content is adjusted to 50%～60%, pH value to 7.5～8 after 1 mix homogeneously,
Obtain substrate；Add 5g CaCO in every kilogram of substrate3, after mixing, load 121 DEG C of sterilizing 20min in fermentation flask, then press
The ratio of mass ratio 10%-15% accesses the bacterium solution that step (1) is obtained, 28 DEG C～30 DEG C, mixing speed 50～70rpm fermentation training
Support 5～7 days, prepared biological prevention and control agent.
Described Duolun lake streptomycete M1-4 microbial inoculum to the main pathogenic fungi causing soybean phytophthora root rot, including Semen sojae atricolor epidemic disease
Mould (P.sojae), Fusarium oxysporum (Fusariumoxysporun) have compared with high inhibition effect, in addition same to other pathogen
Sample is inhibited, for example Herba bromi japonici Fusariumsp (Fusarium avenaceum), gibberella saubinetii
(FusariumGraminearu), Fusarium graminearum (Fusarium graminearum).Duolun lake streptomycete M1-4 is mainly
Antagonism is produced to soybean phytophthora by the secondary metabolite that its fermentation produces.
The biological prevention and control agent preventing and treating soybean phytophthora of the described streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria
The potted plant application test result of root rot shows, biological prevention and control agent is more than 70% to the preventive effect of Soybean Seedling phytophthora root rot, effect
Fruit is preferably.This biological prevention and control agent can effectively prevent and treat soybean diseases, it is to avoid brought series of problems applied by chemical pesticide, has
Beneficial to the No-harmful apple orchard of Semen sojae atricolor, improve soybean quality, reduce Pesticide use and put into, reduce expenses, simultaneously soil protection and ring
Border safety.
Brief description
Fig. 1 is the compares figure of the inhibitory action to phytophthora sojae kaufmann＆gerdemann for the Duolun lake streptomycete M1-4；
Fig. 2 is the design sketch of the inhibitory action to phytophthora sojae kaufmann＆gerdemann for the Duolun lake streptomycete M1-4；
Fig. 3 is the compares figure of the inhibitory action to Fusarium oxysporum for the Duolun lake streptomycete M1-4；
Fig. 4 is the design sketch of the inhibitory action to Fusarium oxysporum for the Duolun lake streptomycete M1-4；
Fig. 5 is the compares figure of the inhibitory action to Fusariumsp for the Duolun lake streptomycete M1-4；
Fig. 6 is the design sketch of the inhibitory action to Fusariumsp for the Duolun lake streptomycete M1-4；
Fig. 7 is the compares figure of the inhibitory action to gibberella saubinetii for the Duolun lake streptomycete M1-4；
Fig. 8 is the design sketch of the inhibitory action to gibberella saubinetii for the Duolun lake streptomycete M1-4；
Fig. 9 is the compares figure of the inhibitory action to Fusarium graminearum for the Duolun lake streptomycete M1-4；
Figure 10 is the design sketch of the inhibitory action to Fusarium graminearum for the Duolun lake streptomycete M1-4；
Figure 11 is the compares figure to the potted plant efficiency test of soybean phytophthora for the Duolun lake streptomycete M1-4；
Figure 12 is the design sketch to the potted plant efficiency test of soybean phytophthora for the Duolun lake streptomycete M1-4；
Figure 13 is Duolun lake streptomycete M1-4 bacterial strain systematic growth tree graph；
Figure 14 is Duolun lake streptomycete M1-4 displaing micro picture；
Figure 15 is Duolun lake streptomycete M1-4 bacterium colony picture.
Specific embodiment
With reference to embodiment and experiment, the invention will be further described：
The streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria of the present invention is Duolun lake streptomycete
(Streptomyces dorenlacusensis) M1-4, this bacterial strain is deposited in China General Microbiological culture presevation management
The heart, preservation address is Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, and preserving number is CGMCC NO.11789.
The screening technique of this Duolun lake streptomycete (Streptomyces dorenlacusensis) M1-4, walks including following
Suddenly：
(1) gather bed mud sample from lake, dry in dark ventilation, become powder with mortar grinder；
(2) carry out separating using dilution method, soil sample is put in the bottle equipped with bead and sterilized water, concussion, after standing, dilute
Release, preparation mass concentration is 10-3、10-4、10-5Diluent, draws diluent and is added drop-wise to addition cycloheximide (50 μ g/ml) and naphthalene
On the Gause I culture medium flat plate of pyridine keto acid (40 μ g/ml), coating uniformly, culture dish is placed on incubator to be inverted culture, growth
28 DEG C of temperature, cultivates 5～7 days, isolates and purifies；
(3) there are the actinomycetes of inhibitory action to soyabean phytophthora using flat board face-off method screening：By the actinomycetes strain of purification, use
Gause I culture medium culturing, actinomycetes are transferred to Radix Dauci Sativae culture medium center, are cultivated 3 days with Φ 9cm culture dish；Greatly
After bean phytophthora is cultivated 4～5 days in Radix Dauci Sativae culture medium, it is cut into Φ 0.5cm mycelia block, is inoculated into the training containing Radix Dauci Sativae culture medium
Foster ware surrounding decussation line position, culture observed measurement antibacterial circle diameter after 7～10 days, selected the obvious unwrapping wire of antagonism
Bacterium, preserves；
(4) the moleculess identification of bacterial strain：The DNA of the bacterial strain that extraction step (3) obtains, using universal primer 27f and 1492r amplification
Its 16S rDNA, the purified sequencing of PCR primer, obtain full length sequence and carry out Phylogenetic Analysis.First in Ezbio Cloud net
Station carries out sequence similarity analysis, finds its wet streptomycete (Streptomes humidus) similarity highest with report,
98.82%.Then utilize MEGA software building phylogenetic tree, see Figure 13.By phylogenetic tree understand this bacterial strain with
Streptomyces humidus in same branch, is not got together with bacterial strains such as Streptomyces edernsis, and relationship is closed
System is nearer.Speculate that this bacterium is streptomyces novel species, and be named as Duolun lake streptomycete (Streptomyces
dorenlacusensis) M1-4.
Duolun lake streptomycete M1-4 strain morphology, cultural characteristic are observed:Perusal streak inoculation is put down in different culture media
The color change of aerial hyphae, substrate mycelium and soluble pigment on plate, the color taking the period of maturation, as the foundation determining kind, is shown in Table
1 and Figure 15.Using ISP3 culture medium culturing one month, there is flourishing branch using scanning microscope TM3030 (5000 ×) mycelia
(see Figure 14), the long oval of spore.
The cultural characteristic of table 1 Duolun lake streptomycete M1-4 bacterial strain
Note：- expression does not produce soluble pigment
The streptomycete M1-4 bacterial strain physiological and biochemical property identification of Duolun lake：Gelatin liquefaction can be made；H can be produced2S；Cellulose hydrolyzes
Experiment shows that it has stronger cellulose degradation ability.NaCl tolerance range is 0～4.0%.PH value tolerance range 4～10,
Optimum pH is 7.28 DEG C of optimum temperature.Carbon source, nitrogen source are shown in Table 2, table 3 using experimental result.
Table 2 Duolun lake streptomycete M1-4 and its sibling specieses utilization of carbon source comparison in difference table
Note：N represents does not have data
Table 3 actinomycetes Duolun lake streptomycete M1-4 and its sibling specieses nitrogen source utilize comparison in difference
Note：N represents does not have data
The streptomycete M1-4 antagonism identification of Duolun lake：Duolun lake streptomycete M1-4 is inoculated in the middle of Radix Dauci Sativae culture medium, 28 DEG C
Culture 3d, inoculates soybean phytophthora, 28 DEG C of culture 7～10d in its surrounding symmetric position on the 4th day, measures bacteriostatic diameter.M1-4 pair
The compares figure of the inhibitory action of soybean phytophthora is shown in Fig. 1, and effect is shown in Fig. 2, and measurement suppression radius is 17.76mm.Same employing is stood facing each other
Method utilizes PDA culture medium to survey Duolun lake streptomycete M1-4 to Fusarium oxysporum (Fusariumoxysporun), Herba bromi japonici Fusariumsp
(Fusarium avenaceum), gibberella saubinetii (FusariumGraminearu), Fusarium graminearum (Fusarium
) etc. graminearum strong inhibition is had to act on.Suppression radius is respectively 14.49mm, 5.56mm, 5.18mm, 15.94mm, its
In, Fig. 3, Fig. 5, Fig. 7, Fig. 9 are compares figure, and Fig. 2, Fig. 4, Fig. 6, Fig. 8, Figure 10 are design sketch.
The biological prevention and control agent of the described streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria is by Duolun lake chain
It is main for being inoculated into soybean cake powder and Testa Tritici after mycete (Streptomyces dorenlacusensis) M1-4 amplification culture
Carry out fermentation culture in the fermentation medium of raw material and obtain spore sum >=1010The biological prevention and control agent of individual/g.
The preparation method of the described biological prevention and control agent of streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria
(1)：
(1) amplification culture of strain：Take Duolun lake streptomycete (Streptomyces dorenlacusensis) M1-4 bacterial strain
Conidium, is inoculated in liquid seed culture medium, and shaking table culture obtains streptomycete liquid seeds, and the condition of shaking table culture is temperature
28 DEG C of degree, rotating speed 180r/min, incubation time 48h；The formula of described liquid seed culture medium is：Yeast extract 4g/L,
Glucose 4g/L, Fructus Hordei Germinatus extract 10g/L, CaCO32g/L, pH value 7.2；
(2) preparation of biological prevention and control agent：By soybean cake powder and Testa Tritici in mass ratio 1：Adjust after 1 mix homogeneously moisture content to 50%,
PH value, to 7.5, obtains substrate；Add 5g CaCO in every kilogram of substrate3, after mixing, load 121 DEG C of sterilizings in fermentation flask
20min, the bacterium solution that then in mass ratio 10% ratio access step (1) is obtained, 28 DEG C DEG C, the fermentation training of mixing speed 50rpm
Support 5 days, prepared biological prevention and control agent.Duolun lake streptomycete M1-4 spore sum 10 in biological prevention and control agent10Described in individual/g to soybean phytophthora
The preparation method (two) of the biological prevention and control agent of the inhibited streptomycete of root rot pathogenic bacteria：
(1) amplification culture of strain：Take Duolun lake streptomycete (Streptomyces dorenlacusensis) M1-4 bacterial strain
Conidium, is inoculated in liquid seed culture medium, and shaking table culture obtains streptomycete liquid seeds, and the condition of shaking table culture is temperature
30 DEG C of degree, rotating speed 180r/min, incubation time 48h；The formula of described liquid seed culture medium is：Yeast extract 4g/L,
Glucose 4g/L, Fructus Hordei Germinatus extract 10g/L, CaCO32g/L, pH value 7.4；
(2) preparation of biological prevention and control agent：By soybean cake powder and Testa Tritici in mass ratio 1：Adjust after 1 mix homogeneously moisture content to 60%,
PH value, to 8, obtains substrate；Add 5g CaCO in every kilogram of substrate3, after mixing, load 121 DEG C of sterilizings in fermentation flask
20min, the bacterium solution that then in mass ratio 15% ratio access step (1) is obtained, 30 DEG C, mixing speed 70rpm fermentation culture 7
My god, prepared biological prevention and control agent.Duolun lake streptomycete M1-4 spore sum 10 in biological prevention and control agent10Individual/g.
The preparation method of the described biological prevention and control agent of streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria
(3)：
(1) amplification culture of strain：Take Duolun lake streptomycete (Streptomyces dorenlacusensis) M1-4 bacterial strain
Conidium, is inoculated in liquid seed culture medium, and shaking table culture obtains streptomycete liquid seeds, and the condition of shaking table culture is temperature
29 DEG C of degree, rotating speed 180r/min, incubation time 48h；The formula of described liquid seed culture medium is：Yeast extract 4g/L,
Glucose 4g/L, Fructus Hordei Germinatus extract 10g/L, CaCO32g/L, pH value 7.3；
(2) preparation of biological prevention and control agent：By soybean cake powder and Testa Tritici in mass ratio 1：Adjust after 1 mix homogeneously moisture content to 55%,
PH value, to 7.8, obtains substrate；Add 5g CaCO in every kilogram of substrate3, after mixing, load 121 DEG C of sterilizings in fermentation flask
20min, the bacterium solution that then in mass ratio 12% ratio access step (1) is obtained, 29 DEG C, mixing speed 60rpm fermentation culture 6
My god, prepared biological prevention and control agent.Duolun lake streptomycete M1-4 spore sum 10 in biological prevention and control agent10Individual/g.
The potted plant efficiency test of Duolun lake streptomycete M1-4 soybean phytophthora root rot biological prevention and control agent：
Investigate the ability of Duolun lake streptomycete M1-4 biological preparation prevention and control soybean phytophthora root rot soil-borne disease.EXPERIMENTAL DESIGN compares
(test soil, CK) and treatment group (test soil+biological prevention and control agent).Field soil accesses No. 1 biological strain of phytophthora sojae kaufmann＆gerdemann
As experimental control soil.Soyabean phytophthora, after solid culture 5d, accesses in V8 fluid medium, the lower 28 DEG C of trainings of dark condition
Support 5 days, pour out fluid medium, mycelia is weighed, access by the amount of 10g/ kilogram of soil.Treatment group is with being connected to soyabean phytophthora
Soil, accesses Duolun lake streptomycete M1-4 biological prevention and control agent in 10g/ kilogram of soil ratio.To compare and process dress basin respectively, every basin
Load 500g soil, eastern agriculture 42 seed (No. 1 biological strain sensitivity) of sowing 10, keep a full stand of seedings 6 plants, be repeated 4 times.Raw in basin alms bowl
Long to 35 days sampling statistics occurring degrees, calculate disease index, measure the total fresh weight of plant, root fresh weight investigates growth-promoting effect, sees figure
11 and Figure 12.
The streptomycete M1-4 process of potted plant applying Duolun lake, shows preferable prevention effect, its sickness rate prevention effect
More than 70%, apply Duolun lake streptomycete M1-4 biological prevention and control agent makes soybean stem leaf fresh weight, root fresh weight increased 120% respectively simultaneously
With 184%, there are obvious growth-promoting functions, be shown in Table 4.
The each impact processing to soybean phytophthora root rot prevention effect, Semen sojae atricolor growth of table 4 greenhouse pot culture experiment
Note:Soybean phytophthora root rot occurring degree is divided into 6 grades.Do not fall ill for root system for 0 grade；1 grade is root system sickness rate≤30%, leaf
Piece is normal；2 grades is 30%<Root system sickness rate≤60%, blade is normal；3 grades is 60%<Root system sickness rate≤80%；Blade becomes
Huang, 4 grades is root system sickness rate more than 80%, and blade is withered, and 5 grades is that whole strain is dead, and blade dries up.
Disease index and prevention effect calculate respectively according to the following formula:Disease index=[(sick level strain number x representative value)/
(total strain number x highest disease level representative value)] x100.Prevention effect=[(comparison disease index-process disease index)/comparison state of an illness
Index] x100%.
Duolun lake streptomycete M1-4 soybean phytophthora root rot biological prevention and control agent field test：
Duolun lake streptomycete M1-4 biological prevention and control agent field controling test is carried out in Semen sojae atricolor second crop soil and academy of agricultural sciences breeding base, test
No the fermentation soybean cake powder of biological prevention and control agent and Testa Tritici are applied in comparison, process and apply Duolun lake streptomycete M1-4, and amount of application is
6000kg/ha, using eastern agriculture 42, cell sets 5 meter of 4 ridge, row spacing 65cm to soybean varieties, often processes 4 repetitions.Semen sojae atricolor V3, V5,
The data such as R1, R3, R5 and period of maturation Investigate incidence, biological yield, statistics finds that M1-4 biological prevention and control agent can significantly reduce Semen sojae atricolor
The sickness rate of phytophthora root rot and raising soybean yields, are shown in Table 5.
Table 5 field is respectively processed to soybean phytophthora root rot prevention effect
The present invention prevents and treats this key issue and sets about from carrying out soybean diseases using biological pesticide antagonistic microbe, develops one kind
The Duolun lake streptomycete M1-4 biological prevention and control agent of soybean phytophthora root rot generation can be significantly reduced, control in field rate reaches more than 60%.
The Antagonistic Actinomycetes that the mechanism of action of this biological prevention and control agent is in this preparation have preferable antagonism, produce antagonistic substance,
The growth of suppression soybean rhizosphere pathogenic microorganism.On the other hand, ferment this biological prevention and control agent substrate be Testa Tritici, soybean cake powder etc. easily
Obtain and cheap raw material, greatly reduce the cost of biological prevention and control agent, preventive effect is also preferable.
Claims (4)
1. a kind of streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria, this bacterial strain is Duolun lake streptomycete
(Streptomyces dorenlacusensis) M1-4, this bacterial strain is deposited in China General Microbiological culture presevation management
The heart, preserving number is CGMCC NO.11789.
2. the biological prevention and control agent of streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria as claimed in claim 1, its
It is characterised by：This biological prevention and control agent is by the Duolun lake streptomycete (Streptomyces described in claim 1
Dorenlacusensis) it is inoculated into after M1-4 amplification culture in the fermentation medium with soybean cake powder and Testa Tritici as primary raw material
Carry out fermentation culture and obtain spore sum >=1010The biological prevention and control agent of individual/g.
3. the preparation of the biological prevention and control agent of streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria described in claim 2
Method it is characterised in that：This preparation method comprises the following steps：
(1) amplification culture of strain：Take Duolun lake streptomycete (Streptomyces dorenlacusensis) M1-4 bacterial strain
Conidium, is inoculated in liquid seed culture medium, and shaking table culture obtains streptomycete liquid seeds, and the condition of shaking table culture is temperature
28～30 DEG C of degree, rotating speed 180r/min, incubation time 48h；The formula of described liquid seed culture medium is：Yeast extract
4g/L, glucose 4g/L, Fructus Hordei Germinatus extract 10g/L, CaCO32g/L, pH value 7.2～7.4；
(2) preparation of biological prevention and control agent：By soybean cake powder and Testa Tritici in mass ratio 1：Adjust after 1 mix homogeneously moisture content to 50%～
60%th, pH value, to 7.5～8, obtains substrate；Add 5g CaCO in every kilogram of substrate3, after mixing, load 121 in fermentation flask
DEG C sterilizing 20min, then in mass ratio 10%-15% ratio access step (1) be obtained bacterium solution, 28 DEG C～30 DEG C, stirring
Speed 50～70rpm fermentation culture 5～7 days, prepared biological prevention and control agent.
4. the preparation of the biological prevention and control agent of streptomycete inhibited to soybean phytophthora root rot pathogenic bacteria described in claim 3
Method it is characterised in that：This preparation method comprises the following steps：
(1) amplification culture of strain：Take Duolun lake streptomycete (Streptomyces dorenlacusensis) M1-4 bacterial strain
Conidium, is inoculated in liquid seed culture medium, and shaking table culture obtains streptomycete liquid seeds, and the condition of shaking table culture is temperature
29 DEG C of degree, rotating speed 180r/min, incubation time 48h；The formula of described liquid seed culture medium is：Yeast extract 4g/L,
Glucose 4g/L, Fructus Hordei Germinatus extract 10g/L, CaCO32g/L, pH value 7.3；
(2) preparation of biological prevention and control agent：By soybean cake powder and Testa Tritici in mass ratio 1：Adjust after 1 mix homogeneously moisture content to 55%,
PH value, to 7.8, obtains substrate；Add 5g CaCO in every kilogram of substrate3, after mixing, load 121 DEG C of sterilizings in fermentation flask
20min, the bacterium solution that then in mass ratio 12% ratio access step (1) is obtained, 29 DEG C, mixing speed 60rpm fermentation culture 6
My god, prepared biological prevention and control agent.
Priority Applications (1)
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434488A (en) * | 2016-11-17 | 2017-02-22 | 黑龙江省农业科学院耕作栽培研究所 | Entomopathogenic nematode symbiotic bacteria with soybean phytophthora root rot inhibiting function and application |
| CN108587981A (en) * | 2018-05-30 | 2018-09-28 | 西南大学 | Multi-functional molten algae streptomycete Streptomyces amritsarensis and its application |
| CN109042015A (en) * | 2018-08-02 | 2018-12-21 | 安徽昌发实业有限公司 | A kind of braiding control method of palm plant phytophthora root rot |
| CN109370959A (en) * | 2018-12-21 | 2019-02-22 | 齐齐哈尔大学 | A kind of rhizosphere bacteria and application thereof preventing and treating the late blight of potato |
| CN112021335A (en) * | 2020-09-10 | 2020-12-04 | 重庆大学 | Method for preventing and treating plant oomycete diseases by combining biological pesticide and chemical pesticide |
| CN112725250A (en) * | 2021-03-10 | 2021-04-30 | 扬州大学 | Streptomyces lavendulae strain 2-1-2F-1 and application of biomass thereof in prevention and treatment of vegetable soil-borne diseases |
| CN116555043A (en) * | 2023-03-12 | 2023-08-08 | 西南大学 | Streptomyces albus capable of preventing and treating pepper epidemic disease and application thereof |
| CN118028143A (en) * | 2023-12-29 | 2024-05-14 | 浙江大学 | Streptomyces albus strain and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103131646A (en) * | 2011-11-29 | 2013-06-05 | 中国科学院沈阳应用生态研究所 | Germ bacteria agent and preparation method and application thereof |
| CN103981126A (en) * | 2014-04-16 | 2014-08-13 | 中国中化股份有限公司 | Microbial agent, preparation method and application thereof |
| CN103992967A (en) * | 2014-04-16 | 2014-08-20 | 中国中化股份有限公司 | Streptomyces albidoflavus and its application in plant disease control |
| CN105385623A (en) * | 2015-11-20 | 2016-03-09 | 江苏省农业科学院 | Actinomycete strain JSX-1 for preventing and curing phytophthora sojae and fungicide thereof |
-
2016
- 2016-07-28 CN CN201610605503.0A patent/CN106399152B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103131646A (en) * | 2011-11-29 | 2013-06-05 | 中国科学院沈阳应用生态研究所 | Germ bacteria agent and preparation method and application thereof |
| CN103981126A (en) * | 2014-04-16 | 2014-08-13 | 中国中化股份有限公司 | Microbial agent, preparation method and application thereof |
| CN103992967A (en) * | 2014-04-16 | 2014-08-20 | 中国中化股份有限公司 | Streptomyces albidoflavus and its application in plant disease control |
| CN105385623A (en) * | 2015-11-20 | 2016-03-09 | 江苏省农业科学院 | Actinomycete strain JSX-1 for preventing and curing phytophthora sojae and fungicide thereof |
Cited By (13)
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| CN106434488A (en) * | 2016-11-17 | 2017-02-22 | 黑龙江省农业科学院耕作栽培研究所 | Entomopathogenic nematode symbiotic bacteria with soybean phytophthora root rot inhibiting function and application |
| CN108587981A (en) * | 2018-05-30 | 2018-09-28 | 西南大学 | Multi-functional molten algae streptomycete Streptomyces amritsarensis and its application |
| CN108587981B (en) * | 2018-05-30 | 2021-10-08 | 西南大学 | Streptomyces amyrosarensis and application thereof |
| CN109042015A (en) * | 2018-08-02 | 2018-12-21 | 安徽昌发实业有限公司 | A kind of braiding control method of palm plant phytophthora root rot |
| CN109042015B (en) * | 2018-08-02 | 2020-12-25 | 安徽昌发实业有限公司 | Method for preventing and controlling phytophthora root rot of palm plants for weaving |
| CN109370959B (en) * | 2018-12-21 | 2021-07-20 | 齐齐哈尔大学 | Rhizosphere bacterium for preventing and treating potato late blight and application thereof |
| CN109370959A (en) * | 2018-12-21 | 2019-02-22 | 齐齐哈尔大学 | A kind of rhizosphere bacteria and application thereof preventing and treating the late blight of potato |
| CN112021335B (en) * | 2020-09-10 | 2022-01-28 | 重庆大学 | Method for preventing and treating plant oomycete diseases by combining biological pesticide and chemical pesticide |
| CN112021335A (en) * | 2020-09-10 | 2020-12-04 | 重庆大学 | Method for preventing and treating plant oomycete diseases by combining biological pesticide and chemical pesticide |
| CN112725250A (en) * | 2021-03-10 | 2021-04-30 | 扬州大学 | Streptomyces lavendulae strain 2-1-2F-1 and application of biomass thereof in prevention and treatment of vegetable soil-borne diseases |
| CN116555043A (en) * | 2023-03-12 | 2023-08-08 | 西南大学 | Streptomyces albus capable of preventing and treating pepper epidemic disease and application thereof |
| CN118028143A (en) * | 2023-12-29 | 2024-05-14 | 浙江大学 | Streptomyces albus strain and application thereof |
| CN118028143B (en) * | 2023-12-29 | 2024-10-11 | 浙江大学 | Streptomyces albus strain and application thereof |
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