CN108753908A - A kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain - Google Patents
A kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain Download PDFInfo
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Abstract
A kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain, actinomyces and Phytophthora nicotianae are seeded to the culture dish equipped with PDA culture medium, the actinomycetes strain that there is antagonism to Phytophthora nicotianae is filtered out by measuring antibacterial band, there are four around the rectangular bars region of its central distribution for setting in the culture dish, actinomyces after activation are applied in four rectangular bars regions, and before being inoculated with Phytophthora nicotianae, the culture dish for smearing actinomyces is put into insulating box and is cultivated 2-4 days, then the Phytophthora nicotianae after inoculation activates at culture dish center again, antibacterial band is measured after being cultivated in insulating box.Actinomyces are inoculated with before inoculation Phytophthora nicotianae in advance and cultivate certain time, be conducive to the formation and release of actinomycetes strain antibiotic, the phenomenon that Phytophthora nicotianae antagonism, can adequately be showed, increase the validity and specific aim of Screening Antagonistic Actinomycetes, the cost for repeating experiment has been saved, has been improved work efficiency.
Description
Technical field
The present invention relates to a kind of bacterial screening method, specifically a kind of quickly screening Phytophthora nicotianae Antagonistic Actinomycetes bacterium
The method of strain.
Background technology
Phytophthora nicotianae (（Phytophthora parasitica var. nicotianae (Breda de Hean)
Tuke) infect balck shank caused by tobacco and be distributed widely in World tobacco growing area, China Henan, Shandong, Anhui, Yunnan,
The cigarette districts such as Guizhou, Guangdong, Fujian, Hunan generally occur, endanger seriously.
Since the generally application of chemical agent makes the remaining increase of pesticide in tobacco leaf and vega soil, the anti-of disease is resulted in
Pharmacological property enhances and rampant again, the not only sustainable development of influence tobacco, while jeopardizing human health, destroys the ecological balance.Therefore,
It is very necessary for tobacco sustainable development to widely popularize biological control.Biocontrol microorganism refers to that those can be used for preventing and plants
The beneficial microbe of object disease.Currently, with the continuous deepening of research, it is comprehensive that the application of Biocontrol microorganism has become plant pest
The important means administered is closed, while being also to preserve the ecological environment, realizes the strong guarantee of agricultural sustainable development.People pass through certainly
The microbial pesticide of right strain breeding thereof exploitation, which has had, to be quite widely applied, and important work is played to the development of agricultural production
With.
Actinomyces (Actinomycetes) belong to prokaryotes, it is distributed widely in nature, type is various, metabolic function
It is different, it is a kind of microbial resources for having extensive use.Actinomyces are a series of important lifes of bioactive substances such as antibiotic
Production person, in the up to ten thousand kinds of antibiotic reported in the world at present, 70% or more is generated by actinomyces.Actinomyces are people's research
Earliest Biocontrol microorganism has antagonism to a variety of pathogens of various crop.Actinomyces are found from Cohn (1872) extremely
The present has 69 1687 kinds of actinomyces of category and is successively reported that there are many shaped preparations to be applied in production, such as jinggangmycin, interior
Element, 768, S-921, pesticide corrosion 120 etc. are treated, it is streptomycete using actinomyces apply in biocontrol of plant disease at most
(StreptomycesSpp.) and its mutation, in addition micromonospora (MicromonosporaSp.) also there is preferable prevention
Effect.
Actinomyces mainly by direct repression to germ or pass through and enhance plant disease-resistant effect and play biocontrol effect.
There are antibiosis, Competition and hyperparasitism to the direct effect of germ, these effects can inhibit pathogen growth, have
A little biocontrol microorganisms can even inhibit the generation of germ virulence factor, to reduce the pathogenicity of germ；Actinomyces can also excite
Plant improves disease resistance, increases antibacterial material synthesis, and inhibits growth, the breeding of pathogen with this, reaches the mesh for mitigating disease
's.
In recent years, the research that tobacco diseases are prevented using Antagonistic Actinomycetes is more and more, and antagonism unwrapping wire is detached from tobacco
Bacterium and tobacco endogeny rayungus are always research hotspot.Traditional opposite culture method screening is suitable for similar similar in growth rate
Face-off between bacterial strain and bacterial strain, such as fungi and fungi, bacterium and bacterium, actinomyces and actinomyces, are not suitable for growth speed
Spend the face-off between fast bacterial strain and the slower bacterial strain of the speed of growth, such as actinomyces and fungi, actinomyces and bacterium.
Since actinomyces type is various, growth speed is again very slow compared with cause of disease Phytophthora nicotianae bacterial strain, conventional
In opposite culture method, the slow bacterial strain of growth rate is easy to be covered by the fast bacterial strain of growth rate, can not observe face-off
Phenomenon, this is that biological existence competition for space is determined.Therefore, conventional opposite culture method is unsuitable for the sieve of antagonistic strain
Choosing.
Invention content
The technical problem to be solved by the present invention is to overcome drawbacks described above, providing one kind can be fast and effectively to from soil
The method that a large amount of actinomycetes strains of acquisition are measured the antagonism of Phytophthora nicotianae is detached in earth.
Used technical solution is the present invention to solve above-mentioned technical problem：One kind quickly put by screening Phytophthora nicotianae antagonism
Actinomyces and Phytophthora nicotianae are seeded to the culture dish equipped with PDA culture medium by the method for line bacteria strain, are sieved by measuring antibacterial band
The actinomycetes strain that there is antagonism to Phytophthora nicotianae is selected, there are four around its central distribution for setting in the culture dish
Rectangular bars region, the actinomyces after activation are applied in four rectangular bars regions, and before being inoculated with Phytophthora nicotianae,
The culture dish for smearing actinomyces is put into insulating box and is cultivated 2-4 days, the cigarette after then inoculation activates at culture dish center again
Careless phytophthora measures antibacterial band after being cultivated in insulating box.
Further, four rectangular bars regions are uniformly distributed around culture dish center, and four rectangular bars
The same circumference is cut in outside region.
Further, the length of a diameter of 9cm of the culture dish, rectangular bars region are 2cm, width 4mm, length
The vertical range at rectangular banded zone and culture dish center is 3cm.
Further, it is seeded to a diameter of 6mm of Phytophthora nicotianae at culture dish center.
Further, it while the Phytophthora nicotianae after activation is seeded to the culture dish, is also provided at another
Phytophthora nicotianae after the center access activation of the culture dish of PDA culture medium goes out antibacterial band as reference by measurement of comparison.
Further, it is 25 DEG C that cultivation temperature is controlled in the insulating box.
The actinomycetes strain obtained is detached before screening, first cross on Gause I culture medium activation culture 3-5d.
Further, then Phytophthora nicotianae bacterial strain is used further to screening actinomyces first at PDA culture medium activation culture 2-4d.
Further, the ingredient of the Gause I culture medium is：Per 20 g of 1000mL culture mediums soluble-containing starch,
KNO3 1 g、NaCl 0.5 g、K2HPO4 0.5g、FeSO4·7H2O 0.01 g、MgSO4·7H20.5 g of O, agar 15 ~ 20
G, surplus are water, and Medium's PH Value is 7.2 ~ 7.4.
Further, the ingredient of the PDA culture medium is：Per 1000mL culture mediums 200g containing potato, glucose 20g,
Agar 20g, surplus are water, Medium's PH Value 7.0.
The beneficial effects of the invention are as follows：
1, the actinomycetes strain that will isolate and purify acquisition early period, before being inoculated with Phytophthora nicotianae, positioning in advance is seeded in PDA plate
On culture dish, 25 DEG C of constant incubator culture 2-4d are placed in, the formation and release of actinomycetes strain antibiotic are conducive to, if
The bacterial strain has antagonism to Phytophthora nicotianae, and it is abundant to form with the antibiotic of release energy the phenomenon that Phytophthora nicotianae antagonism
Show, increase the validity and specific aim of Screening Antagonistic Actinomycetes, solve that the actinomyces speed of growth is slow, Bu Nengyu
The technical issues of Phytophthora nicotianae opposite culture, has saved the cost for repeating experiment, has improved work efficiency.
2, four rectangular bars regions on culture dish are set, form the face-off of four direction with the Phytophthora nicotianae at center,
Multiple measurement results can be once obtained, the group number of experiment is reduced, reduce operating quantity, shorten the screening period.
3, the method that the present invention quickly screens Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain, is not only suitable for actinomyces and tobacco epidemic disease
Mould opposite culture, and actinomyces are also suitable for sickle-like bacteria（Fusariumspp.）, pythium spp（Pythiumspp.）,
Rhizoctonia（Rhizoctoniaspp.）The opposite culture of equal disease fungus.
Description of the drawings
Fig. 1 is rectangular bars region installation position and the inoculation position embodiment signal of center Phytophthora nicotianae in culture dish
Figure.
Fig. 2 is the embodiment schematic diagram of the culture dish center inoculation Phytophthora nicotianae of reference.
It is marked in figure：1, screening group culture dish, 2, rectangular bars region, 3, screening group Phytophthora nicotianae, 4, with reference to tissue culture
Support ware, 5, reference group Phytophthora nicotianae.
Specific implementation mode
Clear, complete explanation is carried out to technical scheme of the present invention below in conjunction with the drawings and the specific embodiments.Below
Particular content listed by embodiment is not limited to skill necessary to the technical solution technical problems to be solved of claim record
Art feature.Meanwhile described enumerate is a part that embodiment is only the present invention, rather than whole embodiments.
The method that the present invention quickly screens Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain, first as shown in Figure 1, being arranged in culture dish
There are four around the rectangular bars region of its central distribution, four rectangular bars regions may be used at the culture dish back side stroke
The modes such as line, placement marker are arranged, and four rectangular bars regions are uniformly distributed around culture dish center, and four rectangular bars
The same circumference is cut in outside region.The PDA culture medium after melting is poured into culture dish, makes PDA plate, it is spare.
The size of rectangular bars region is determined according to the size of culture dish, by taking the culture dish of 9cm diameters as an example（Similarly hereinafter）,
The setting length of rectangular bars region is 2cm, width 4mm, the vertical range of rectangular bars region and culture dish center
For 3cm.
Since Antagonistic Actinomycetes are to the antagonism major embodiment of phytopathogen secondarily grade metabolite, that is, antibiotic
Pathogen is directly inhibited and lethal effect, the influence that pathogen cell and macromolecular substances are synthesized by antibiotic（Table
Inhibit the synthesis of protein, nucleic acid and plasma membrane, or the metabolic system of interference pathogen now）, to inhibit growth.Therefore,
The present invention is in order to fully show inhibiting effect of the actinomyces to Phytophthora nicotianae, and before being inoculated with Phytophthora nicotianae, inoculation in advance is put
Line bacterium is simultaneously cultivated certain time, and the formation and release of actinomycetes strain antibiotic are conducive to.Specific method is：In inoculation tobacco epidemic disease
Before mould, first the actinomyces after activation are uniformly applied in four rectangular bars regions, pay attention to not exceeding length when coating
Rectangular lace boundary line.Then the culture dish for smearing actinomyces is put into 25 DEG C of insulating boxs and is cultivated 2-4 days.
It is inoculated with actinomyces in advance and cultivates after a certain period of time, the Phytophthora nicotianae after the inoculation activation of culture dish center, in perseverance
Antibacterial band is measured after being cultivated in incubator, the actinomyces bacterium that there is antagonism to Phytophthora nicotianae is filtered out by measuring antibacterial band
Strain.
It is same at another while the Phytophthora nicotianae of activation is seeded to the culture dish for the ease of measuring antibacterial band
Phytophthora nicotianae after culture dish center access activation of the sample equipped with PDA culture medium goes out antibacterial band as reference by measurement of comparison.
For example, inoculation Phytophthora nicotianae is as screening group in the culture dish of inoculation actinomyces and culture 2-4 days, another
A culture dish center for being also provided with PDA is inoculated with Phytophthora nicotianae as reference group.The Phytophthora nicotianae diameter of inoculation is 6mm.
The growing state for observing Phytophthora nicotianae and two kinds of bacterial strains of actinomyces after inoculation daily, when reference group Phytophthora nicotianae bacterium colony
When diameter is grown to 5-7cm, the presence or absence of observation antibacterial band of screening group measures antibacterial band diameter（Width）, true according to antibacterial band size
Determine bacteriostasis rate, and preliminary screening provides the defence line bacteria strain of antagonism.
Wherein, bacteriostasis rate=（Compare colony diameter-processing colony diameter）/（Compare colony diameter-bacteria cake diameter）×
100%。
Due on culture dish be provided with four smearing actinomyces rectangular bars regions, can be formed on four direction and
The face-off of the Phytophthora nicotianae at center, testing result is more accurate, once obtains multiple measurement results, reduces experimental group number.
Above-described actinomyces and Phytophthora nicotianae are being inoculated with for all being realized before screening by activation process, specific side
Method is：The actinomycetes strain obtained is detached before screening, first cross on Gause I culture medium activation culture 3-5d.Tobacco
Mtr mutant first in PDA culture medium activation culture 2-4d, is then used further to screening actinomyces.
The ingredient of the Gause I culture medium is：Per 1000mL culture mediums soluble-containing starch 20 g, KNO3 1 g、
NaCl 0.5 g、K2HPO4 0.5g、FeSO4·7H2O 0.01 g、MgSO4·7H20.5 g of O, 15 ~ 20 g of agar, surplus are
Water, Medium's PH Value are 7.2 ~ 7.4.The ingredient of the PDA culture medium is：Per 1000mL culture mediums 200g containing potato, grape
Sugared 20g, agar 20g, surplus are water, Medium's PH Value 7.0.
Embodiment 1
A kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain, operating procedure are：
Step 1：Actinomycetes strain that separation obtains is crossed activation culture 4d on Gause I culture medium；Phytophthora nicotianae Breda
Strain is in PDA culture medium activation culture 3d；It is spare.
Step 2：2 cm wide 4 long, 4 mm parallel two-by-two are drawn in the culture dish bottom outer of sterilized diameter 9cm
Rectangular strip, with culture dish center vertical range be 3 cm（Fig. 1）, the PDA culture medium after melting is poured into, it is flat to make PDA
Plate, it is spare.
Step 3：The actinomyces that step 1 activates are uniformly coated on to the corresponding rectangle of PDA plate of step 2 preparation
In band, pays attention to not exceeding rectangular strip boundary line when coating, be placed in 25 DEG C of constant incubator culture 3d.
Step 4：The Phytophthora nicotianae of step 1 activation is got into bacteria cake with the card punch of diameter 6mm, is transferred in step 2 institute
The PDA plate center stated, meanwhile, using Phytophthora nicotianae of only transferring as reference, the Phytophthora nicotianae bacteria cake switching of diameter 6mm is flat in PDA
Plate center（Fig. 2）；Two kinds of PDA plates are placed in 25 DEG C of constant incubator cultures.
Step 5：The growing state for observing Phytophthora nicotianae and two kinds of bacterial strains of actinomyces after inoculation daily, when the tobacco of reference
When phytophthora colony diameter is grown to 7cm, whether there is or not antibacterial bands in the culture dish of observation screening, measure antibacterial band diameter（Width）, really
Determine bacteriostasis rate, preliminary screening provides the defence line bacteria strain of antagonism accordingly.
Embodiment 2
A kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain, operating procedure are：
Step 1：Actinomycetes strain that separation obtains is crossed activation culture 4d on Gause I culture medium；Phytophthora nicotianae Breda
Strain is in PDA culture medium activation culture 4d；It is spare.
Step 2：2 cm wide 4 long, 4 mm parallel two-by-two are drawn in the culture dish bottom outer of sterilized diameter 9cm
Rectangular strip, with culture dish center vertical range be 3 cm（Fig. 1）, the PDA culture medium after melting is poured into, it is flat to make PDA
Plate, it is spare.
Step 3：The actinomyces that step 1 activates are uniformly coated on to the corresponding rectangle of PDA plate of step 2 preparation
In band, pays attention to not exceeding rectangular strip boundary line when coating, be placed in 25 DEG C of constant incubator culture 4d.
Step 4：The Phytophthora nicotianae of step 1 activation is got into bacteria cake with the card punch of diameter 6mm, is transferred in step 2 institute
The PDA plate center stated；Meanwhile using Phytophthora nicotianae of only transferring as reference, the Phytophthora nicotianae bacteria cake switching of diameter 6mm is flat in PDA
Plate center（Fig. 2）；Two kinds of PDA plates are placed in 25 DEG C of constant incubator cultures.
Step 5：The growing state for observing Phytophthora nicotianae and two kinds of bacterial strains of actinomyces after inoculation daily, when the tobacco of reference
When phytophthora colony diameter is grown to 6cm, whether there is or not antibacterial bands in the culture dish of observation screening, measure antibacterial band diameter（Width）, really
Determine bacteriostasis rate, preliminary screening provides the defence line bacteria strain of antagonism accordingly.
Embodiment 3
A kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain, operating procedure are：
Step 1：Actinomycetes strain that separation obtains is crossed activation culture 4d on Gause I culture medium；Phytophthora nicotianae Breda
Strain is in PDA culture medium activation culture 2d；It is spare.
Step 2：2 cm wide 4 long, 4 mm parallel two-by-two are drawn in the culture dish bottom outer of sterilized diameter 9cm
Rectangular strip, with culture dish center vertical range be 3 cm（Fig. 1）, the PDA culture medium after melting is poured into, it is flat to make PDA
Plate, it is spare.
Step 3：The actinomyces that step 1 activates are uniformly coated on to the corresponding rectangle of PDA plate of step 2 preparation
In band, pays attention to not exceeding rectangular strip boundary line when coating, be placed in 25 DEG C of constant incubator culture 2d.
Step 4：The Phytophthora nicotianae of step 1 activation is got into bacteria cake with the card punch of diameter 6mm, is transferred in step 2 institute
The PDA plate center stated；Meanwhile using Phytophthora nicotianae of only transferring as reference, the Phytophthora nicotianae bacteria cake switching of diameter 6mm is flat in PDA
Plate center（Fig. 2）；It is placed in 25 DEG C of constant incubator cultures.
Step 5：The growing state for observing Phytophthora nicotianae and two kinds of bacterial strains of actinomyces after inoculation daily, when the tobacco of reference
When phytophthora colony diameter is grown to 5cm, whether there is or not antibacterial bands in the culture dish of observation screening, measure antibacterial band diameter（Width）, really
Determine bacteriostasis rate, preliminary screening provides the defence line bacteria strain of antagonism accordingly.
Inventor has selected multiple places, tobacco roll into a ball a phase, it is prosperous for a long time and picking time, cigarette is acquired using five point sampling
Grass roots encloses soil sample.Each sampling point randomly selects 5 ~ 6 healthy and strong cigarette strains and first removes away from the side within the scope of 30 cm of plant trunk
The table soil of 0 ~ 5 cm, excavates soil profile with spades, the band root soil sample of 5 ~ 20 cm soil layers is acquired, by collected band root soil sample
After mixing well, retains sample according to quartering, be fitted into polybag after rejecting impurity and take back laboratory；Record sample number into spectrum,
Sampling position, sampling time.The soil sample adopted back is placed on newspaper, is spread out at very thin one layer, as room-dry；In soil sample half
When dry, large clod especially cohesive soil is crumbed, in order to avoid form lump after air-drying completely, it is difficult to it is levigate；Soil sample after air-drying,
Pick plant and animal residues such as root, stem, leaf, polypide, stone etc.；Then the glass mortar grinding disinfected in alcohol, crosses the sieve of 20 mesh
Son saves backup.The separation of soil actinomycete is isolated using Gause I culture medium, the separation of conventional dilution-plate method
496 actinomyces single bacterium colonies obtain 227 plants different of actinomyces of form through being further purified.
227 plants of actinomyces are randomly divided into 16 groups（Table 1, table 2）, these bacterial strains are measured according to the method and step of embodiment 1
To the inhibiting effect of Phytophthora nicotianae, test result（Table 3, table 4）Show there are 47 plants of actinomyces to exist the bacteriostasis rate of Phytophthora nicotianae
50% or more, secondary screening program can be entered and carry out the identification of actinomyces type, for further study, utilization.
1 actinomycetes strain of table is tested Phytophthora nicotianae inhibiting effect short of money and is grouped
2 actinomycetes strain of table is tested Phytophthora nicotianae inhibiting effect short of money and is grouped
Inhibiting effect measurement result of 3 actinomycetes strain of table to Phytophthora nicotianae（Bacteriostasis rate >=50%）
Inhibiting effect measurement result of 4 actinomycetes strain of table to Phytophthora nicotianae（Bacteriostasis rate >=50%）
The present invention not only solves and cannot measure actinomyces to Phytophthora nicotianae antagonism using traditional tablet opposite culture method
The problem of, and greatly improve the specific aim, validity and working efficiency of screening.
The technical concept and its core concept of the present invention are merely used to help understand to the explanation of specific implementation mode above,
Technical solution is described and illustrated although there is used herein specific preferred embodiments, should not be construed as to this hair
The bright limitation of itself.Those skilled in the art, can be to it in form and carefully under the premise of not departing from the technology of the present invention design
It is made a variety of changes on section.These change or replacement readily occurred in should be covered by the protection scope of the present invention.
Claims (10)
1. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain, actinomyces and Phytophthora nicotianae are seeded to and are equipped with
The culture dish of PDA culture medium filters out the actinomycetes strain for having antagonism to Phytophthora nicotianae by measuring antibacterial band, special
Sign is：There are four around the rectangular bars region of its central distribution, the actinomyces after activation apply for setting in the culture dish
It is put in four rectangular bars regions, and before being inoculated with Phytophthora nicotianae, the culture dish for smearing actinomyces is put into constant temperature
It is cultivated in case 2-4 days, the Phytophthora nicotianae after then inoculation activates at culture dish center again measures suppression after being cultivated in insulating box
Cingula.
2. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as described in claim 1, it is characterised in that：Institute
The four rectangular bars regions stated are uniformly distributed around culture dish center, and four rectangular bars regions are circumscribed in the same circle
Week.
3. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as claimed in claim 2, it is characterised in that：Institute
A diameter of 9cm of culture dish is stated, the length of rectangular bars region is 2cm, width 4mm, rectangular bars region and culture
The vertical range at ware center is 3cm.
4. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as described in claim 1 or 3, feature exist
In：It is seeded to a diameter of 6mm of Phytophthora nicotianae at culture dish center.
5. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as described in claim 1, it is characterised in that：?
While Phytophthora nicotianae after activation is seeded to the culture dish, in the culture dish that another is also provided with PDA culture medium
Phytophthora nicotianae after the access activation of center goes out antibacterial band as reference by measurement of comparison.
6. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as described in claim 1, it is characterised in that：Institute
It is 25 DEG C to state and control cultivation temperature in insulating box.
7. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as described in claim 1, it is characterised in that：Point
Actinomycetes strain from acquisition is before screening, and first cross on Gause I culture medium activation culture 3-5d.
8. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as described in claim 1, it is characterised in that：Cigarette
Careless Mtr mutant first in PDA culture medium activation culture 2-4d, is then used further to screening actinomyces.
9. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as claimed in claim 7, it is characterised in that：Institute
The ingredient for stating Gause I culture medium is：Per 1000mL culture mediums soluble-containing starch 20 g, KNO3 1 g、NaCl 0.5 g、
K2HPO4 0.5g、FeSO4·7H2O 0.01 g、MgSO4·7H20.5 g of O, 15 ~ 20 g of agar, surplus are water, medium pH
Value is 7.2 ~ 7.4.
10. a kind of method of quick screening Phytophthora nicotianae Antagonistic Actinomycetes bacterial strain as claimed in claim 8, it is characterised in that：
The ingredient of the PDA culture medium is：Per 1000mL culture mediums 200g containing potato, glucose 20g, agar 20g, surplus is water,
Medium's PH Value is 7.0.
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| CN110229868A (en) * | 2019-06-19 | 2019-09-13 | 河南省农业科学院烟草研究所 | A kind of test method inhibiting Phytophthora nicotianae Breda |
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| CN110229868A (en) * | 2019-06-19 | 2019-09-13 | 河南省农业科学院烟草研究所 | A kind of test method inhibiting Phytophthora nicotianae Breda |
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