CN106399210A - Fermentation method and application of antagonistic bacteria for tobacco alternaria alternate - Google Patents
Fermentation method and application of antagonistic bacteria for tobacco alternaria alternate Download PDFInfo
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- CN106399210A CN106399210A CN201611152844.3A CN201611152844A CN106399210A CN 106399210 A CN106399210 A CN 106399210A CN 201611152844 A CN201611152844 A CN 201611152844A CN 106399210 A CN106399210 A CN 106399210A
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- Prior art keywords
- alternaria alternate
- fermentation process
- fermentation
- antagonistic fungi
- tobacco
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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
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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
Abstract
The invention belongs to the technical field of tobacco plantation, particularly relates to a fermentation method and application of antagonistic bacteria for tobacco alternaria alternate. The method provides a fermentation method of antagonistic bacteria for tobacco alternaria alternate. The method comprises the following steps: transplanting the antagonistic bacteria for tobacco alternaria alternate in a fermentation culture medium, shaking and culturing to obtain the fermented fluid of the antagonistic bacteria for tobacco alternaria alternate, the shaking and fermentation time lasts for 72 hours. An application of the antagonistic bacteria for tobacco alternaria alternate cultured and prepared by the this technical plan is also included. The bacteria inhibition rate is substantially upgraded. The method can be applied to the prevention and treatment of tobacco alternaria alternate.
Description
Technical field
The invention belongs to tobacco planting field, more particularly, to a kind of fermentation process of Alternaria alternate Antagonistic Fungi and its should
With.
Background technology
Tobacco drafts a document eggplant wood, and Solanaceae is annual or limited herbaceos perennial, base portion slightly lignifying.Inflorescence basidixed, circle
Taper, spends more；Capsule ovate or square round shape, length approximates persistent calyx.Summer and autumn yields positive results.It is distributed mainly on South America, south
Sub-, Chinese.For effectively improving tobacco production, improve production capacity, the disease control of tobacco is particularly important.
Alternaria alternate is caused by tobacco brown spot pathogen, is the disease threatening maximum on the tobacco leaf production of the world each tobacco producing region
One of, all there is generation in each cigarette district of China, annual onset area accounts for the 30～35% of China's cultivated area.At present, mainly adopt
With the Prevention Technique based on chemical prevention, but medicament residue risk and the impact of pathogen resistance risk, limit the big of agricultural chemicals
Scale utilizes.Separate brown spot pathogen is had good inhibitory action and prevention effect Antagonistic Fungi be implement biological control
An important process.
Therefore, a kind of fermentation process of Alternaria alternate Antagonistic Fungi and its application are developed, for solving in prior art,
Chemical prevention Alternaria alternate medicament residue risk and the impact of pathogen resistance risk, become those skilled in the art urgently
The problem solving.
Content of the invention
In view of this, the invention provides a kind of fermentation process of Alternaria alternate Antagonistic Fungi and its application, for solving
In prior art, chemical prevention Alternaria alternate medicament residue risk and may lead to pathogen resistance risk affect, realize
The green prevention and control of Alternaria alternate.Alternaria alternate Antagonistic Fungi, training of can fermenting obtained by technical scheme screening that the present invention provides
Educate and obtain Alternaria alternate Antagonistic Fungi substantial amounts of, that bacteriostatic activity is high, for Alternaria alternate biological control play positive
Preventive and therapeutic effect.
The invention provides a kind of fermentation process of Alternaria alternate Antagonistic Fungi, described fermentation process is：Described tobacco is red
Star disease Antagonistic Fungi is inoculated in fermentation medium, and shaken cultivation obtains Alternaria alternate Antagonistic Fungi；
The time of described shaken cultivation is 72h.
Preferably, the inoculum concentration of described Alternaria alternate Antagonistic Fungi is 0.1%～2%.
Preferably, the inoculum concentration of described Alternaria alternate Antagonistic Fungi is 0.5%.
Preferably, the temperature of described shaken cultivation is 25～45 DEG C.
It is highly preferred that the temperature of described shaken cultivation is 30 DEG C.
Preferably, the initial pH of described fermentation medium is 5～9.
It is highly preferred that the initial pH of described fermentation medium is 6.
Preferably, the rotating speed of described shaken cultivation is 130～170r/min.
It is highly preferred that the rotating speed of described shaken cultivation is 150r/min.
Preferably, the carbon source of described fermentation medium is selected from：One of beef extract, mannitol and D-Fructose or many
Kind.
It is highly preferred that the carbon source of described fermentation medium is beef extract.
Preferably, the nitrogen source of described fermentation medium is selected from：One of peptone, yeast extract and tryptone or
Multiple.
It is highly preferred that the nitrogen source of described fermentation medium is peptone.
Preferably, described fermentation process also includes：Separating-purifying；
Described separating-purifying step is carried out after described shaken cultivation.
Present invention also offers the Alternaria alternate that a kind of fermentation process culture including described in any of the above one is obtained
Antagonistic Fungi is in the application in tobacco disease resistance field.
In sum, the invention provides a kind of fermentation process of Alternaria alternate Antagonistic Fungi, described fermentation process is：Institute
State Alternaria alternate Antagonistic Fungi to be inoculated in fermentation medium, shaken cultivation, obtain Alternaria alternate Antagonistic Fungi；Described shaken cultivation
Time be 72h.Present invention also offers a kind of above-mentioned fermentation process cultivates the Alternaria alternate Antagonistic Fungi being obtained resisting in tobacco
The application in sick field.Can obtain after testing, the Alternaria alternate Antagonistic Fungi that the technical scheme cultivation that the present invention provides obtains, bacteriostasis rate
Significantly improve, can be used for the preventing and treating of Alternaria alternate.
Brief description
In order to be illustrated more clearly that the embodiment of the present invention or technical scheme of the prior art, below will be to embodiment or existing
Have technology description in required use accompanying drawing be briefly described it should be apparent that, drawings in the following description be only this
Inventive embodiment, for those of ordinary skill in the art, on the premise of not paying creative work, can also basis
The accompanying drawing providing obtains other accompanying drawings.
In a kind of fermentation process of Alternaria alternate Antagonistic Fungi that Fig. 1 provides for the present invention, time of shaken cultivation for
The impact result of bacteriostasis rate；
In a kind of fermentation process of Alternaria alternate Antagonistic Fungi that Fig. 2 provides for the present invention, inoculum concentration is for bacteriostasis rate
Impact result；
In a kind of fermentation process of Alternaria alternate Antagonistic Fungi that Fig. 3 provides for the present invention, the temperature of shaken cultivation for
The impact result of bacteriostasis rate；
In a kind of fermentation process of Alternaria alternate Antagonistic Fungi that Fig. 4 provides for the present invention, the initial pH of fermentation medium
Impact result for bacteriostasis rate；
In a kind of fermentation process of Alternaria alternate Antagonistic Fungi that Fig. 5 provides for the present invention, the rotating speed of shaken cultivation for
The impact result of bacteriostasis rate；
In a kind of fermentation process of Alternaria alternate Antagonistic Fungi that Fig. 6 provides for the present invention, the carbon source pair of fermentation medium
Impact result in bacteriostasis rate；
In a kind of fermentation process of Alternaria alternate Antagonistic Fungi that Fig. 7 provides for the present invention, the nitrogen source pair of fermentation medium
Impact result in bacteriostasis rate.
Specific embodiment
The invention provides a kind of fermentation process of Alternaria alternate Antagonistic Fungi and its application, for solving prior art
In, chemical prevention Alternaria alternate medicament residue risk and may lead to pathogen resistance risk affect, realize the red star of tobacco
The green prevention and control of disease.The Alternaria alternate Antagonistic Fungi obtained by technical scheme screening that the present invention provides, cultivation of can fermenting obtains greatly
Alternaria alternate Antagonistic Fungi that measure, that bacteriostatic activity is high, positive preventive and therapeutic effect is played in the biological control for Alternaria alternate.
The enforcement it is clear that described will be clearly and completely described to the technical scheme in the embodiment of the present invention below
Example is only a part of embodiment of the present invention, rather than whole embodiments.Based on the embodiment in the present invention, this area is common
The every other embodiment that technical staff is obtained under the premise of not making creative work, broadly falls into the model of present invention protection
Enclose.
In order to the present invention is described in more detail, a kind of Alternaria alternate Antagonistic Fungi of the present invention being provided with reference to embodiment
Fermentation process and its application, be specifically described.
In following embodiments of the present invention, bacteriostatic activity bacteriostasis rate represents, computing formula is：
Bacteriostasis rate (%)=【(ck colony diameter-process colony diameter)/ck colony diameter】* 100%
Embodiment 1
The present embodiment is the specific embodiment of separate tobacco rust Antagonistic Fungi.
1.1 getting the raw materials ready
Tobacco brown spot pathogen：Separated by Jiangxi Prov. Inst. of Tobacco Leaf Science laboratory and preserve.
Pedotheque：Pick up from the health tobacco plant roots of Ruijin City nonyl Tian Town tobacco seed growing area tobacco brown spot pathogen infected area
Border soil.
PDA culture medium：Potato 200g, glucose 20g, agar 18g and deionized water 1000mL are mixed to prepare.
NA culture medium：Beef extract 3g, peptone 10g, NaCl5g, agar 18g and deionized water 1000mL are mixed to prepare.
Gao Shi I culture medium：Soluble starch 20g, KNO31g、K2HPO40.5g、MgSO4·7H2O 0.5g、
NaCl0.05g、FeSO4·7H2O0.01g, agar 18g and deionized water 1000mL are mixed to prepare.
1.2 soil bacterias and actinomycetic separation and purifying
Weigh the ground soil sample of 10g to load in the triangular flask of sterilizing, add 90mL sterilized water, make 1:10 soil is dilute
Release liquid, fully shaking is cultivated 30 minutes in 28 DEG C of shaking tables, is then made into 10-1～10-5Dilution is standby.Take above-mentioned 5 respectively
Concentration soil dilution liquid 100 μ L, is separately added on ready-made NA and Gao Shi I plating medium, applies rod with the glass of sterilizing
Dilution is uniformly coated with, puts into culture in 28 DEG C of insulating boxs, if three repetitions.Observe by the naked eye picking cultural characteristic not
Same single bacterium colony, is saved in standby in 4 DEG C of refrigerators after purification.
1.3 flat board primary dcreening operations and bacteriostasis rate measure
Primary dcreening operation adopts flat board face-off method, to the actinomyces isolating and purifying and bacterium, so that examination disease fungus is target, adopts
With flat board opposite culture method, directly measured with agar plate.Concrete grammar is：PDA plate center inoculate pathogen bacteria cake, four
Week inoculation actinomycetes strain to be measured.In 28～30 DEG C of incubated 4d, see whether that inhibition zone produces.There is inhibition zone,
Measurement inhibition zone radius, calculates bacteriostasis rate.
1.4 primary dcreening operation antagonism fermented liquid bacteriostatic activity secondary screenings
Select the good actinomyces of Antagonism or bacterium in primary dcreening operation, bred with the method for liquid fermentation and culture, measure
The Antagonism to pathogen for the ferment filtrate, its method is：
(1), actinomyces or bacterium access fluid nutrient medium, water bath with thermostatic control shaking table culture, 28 DEG C, 150r/min, culture
72h.
(2), fermentation liquor pretreatment：Take out after actinomyces Liquid Culture 72h, after standing 1h, aseptically filter, obtain
Ferment filtrate.
(3), growth rate method measures Antagonism：Ferment filtrate is taken in 3mL injection culture dish, then by the PDA of fusing
Culture medium (about 15mL) is poured in ware and is mixed with ferment filtrate, and cooling is followed by the bacteria cake of pathogen.Respectively to add sterilized water
And do not connect bacterium liquid fermentation and culture liquid PDA plate be comparison, after 28 DEG C of incubated 3d measure colony diameter, calculate suppression
Rate processed.
The Molecular Identification of 1.5 Antagonistic Fungis
By the bacterium common molecular authentication method that above-mentioned bacteriostasis rate is optimum, the species of identification Antagonistic Fungi.Identified, this is short of money
Antibacterial is bacillus (Bacillus vanillea).
Embodiment 2
The present embodiment be detection in Alternaria alternate Antagonistic Fungi, that is, in the fermentation process of bacillus, shaken cultivation when
Between for Antagonistic Fungi bacteriostasis rate impact specific embodiment.
With 1% inoculum concentration, antagonism bacteria culture fluid is inoculated in fermentation basal medium, in 28 DEG C, under 150r/min rotating speed
Shaken cultivation 24,48,72,96,120 detects the impact to bacterial strain bacteriostatic activity for the different shaken cultivation with after 144h, determines the most suitable
The shaken cultivation time.
Experimental results refer to Fig. 1.Can be obtained by Fig. 1, the impact that shaken cultivation produces fungistatic effect to zymotic fluid is relatively
Greatly, bacteriostatic activity assumes downward trend after first rising.During shaken cultivation 24～72h bacteriostasis rate with incubation time increase and
It is gradually increased, when fermenting 72h, preferably, bacteriostasis rate reaches maximum to the fungistatic effect of bacterial strain, is 77.72%.Therefore, shaken cultivation
72h to 7 kinds of fermentation times of bacteriostasis rate and other of brown spot pathogen compared with, fungistatic effect is optimal.
Embodiment 3
The present embodiment is detection in Alternaria alternate Antagonistic Fungi, that is, in the fermentation process of bacillus, the temperature of shaken cultivation
Degree is for the specific embodiment of Antagonistic Fungi bacteriostasis rate impact.
Under 1% inoculum concentration, 150r/min speed conditions, from the most suitable shaken cultivation time fixed in embodiment 2,
Fermented with 20 DEG C, 25 DEG C, 30 DEG C, 35 DEG C, 40 DEG C and 45 DEG C of 6 cultivation temperature respectively, detection different temperatures is to Antagonistic Fungi
The impact of bacteriostatic activity, determines the most suitable shaken cultivation temperature of bacterial strain.
Experimental results refer to Fig. 3.Can be obtained by Fig. 3, in the range of 25～45 DEG C, the bacteriostasis rate of bacterial strain is all presented on
One good bacteriostasis；Wherein, the bacteriostatic activity highest when 30 DEG C, inhibiting rate is 53.08%.Therefore, optimum shake
Swing cultivation temperature and be 30 DEG C.
Embodiment 4
The present embodiment is detection in Alternaria alternate Antagonistic Fungi, that is, in the fermentation process of bacillus, fermentation medium
The specific embodiment that initial pH affects for Antagonistic Fungi bacteriostasis rate.
Under conditions of 1% inoculum concentration and 150r/min rotating speed, initial pH value of medium is respectively set to 5,6,7,8,9
With 6 different gradients of 10 grade, fermented from fixed the most suitable fermentation time and temperature, the different pH value of detection is to Antagonistic Fungi
The impact of bacteriostatic activity, determines the most suitable initial pH value of fermentation medium.
Experimental results refer to Fig. 4.Can be obtained by Fig. 4, the initial pH of culture medium be 5～9 in the range of, bacterial strain antibacterial
Rate is all presented on a good bacteriostasis；Wherein, the bacteriostatic activity highest when culture medium is initially 7.Therefore, optimum
Culture medium is initially 7.
Embodiment 5
The present embodiment is to detect in Alternaria alternate Antagonistic Fungi, and that is, in the fermentation process of bacillus, inoculum concentration is for short of money
The specific embodiment of antibacterial bacteriostatic rate impact.
Under conditions of 150r/min rotating speed, respectively with 0.05%, 0.1%, 0.5%, 1.0%, 1.5% and 2.0%
Antagonism bacteria culture fluid is inoculated in basal medium inoculum concentration, from fixed the most suitable fermentation time, temperature and initial pH
Value is fermented, and detects the impact to Antagonistic Fungi bacteriostatic activity for the different vaccination amount, determines the most suitable inoculum concentration.
Experimental results refer to Fig. 2.Can be obtained by Fig. 2, be the suppression of bacterial strain in the range of 0.1%～2% in inoculum concentration
Bacterium rate is all presented on a good bacteriostasis；Wherein, the bacteriostatic activity highest when inoculum concentration is 0.5%.Therefore, optimum
Inoculum concentration be 0.5%.
Embodiment 6
The present embodiment be detection in Alternaria alternate Antagonistic Fungi, that is, in the fermentation process of bacillus, shaken cultivation turn
Speed is for the specific embodiment of Antagonistic Fungi bacteriostasis rate impact.
The shaking speed of setting shaken cultivation is respectively 100r/min, 130r/min, 150r/min, 170r/min, 190r/
Min and 210r/min amounts to 6 different rotating speeds, and in the fixed the most suitable fermentation time of embodiment 2～5, temperature, initial pH value
Fermented with inoculum concentration, detect the impact to Antagonistic Fungi bacteriostatic activity for the different rotating speeds, determine optimum speed.
Experimental results refer to Fig. 5.Can be obtained by Fig. 5, be 130～170r/min's in the shaking speed of shaken cultivation
In the range of, the bacteriostasis rate of bacterial strain is all presented on a good bacteriostasis；Wherein, the shaking speed in shaken cultivation is
Bacteriostatic activity highest during 150r/min.Therefore, the shaking speed of optimum shaken cultivation is 150r/min.
Embodiment 7
The present embodiment is detection in Alternaria alternate Antagonistic Fungi, that is, in the fermentation process of bacillus, fermentation medium
The specific embodiment that carbon source affects for Antagonistic Fungi bacteriostasis rate.
Carbon source is the necessary class important nutrient of growth of microorganism, and the carbohydrate in culture medium is not only micro- life
The main source of thing vital movement energy, and be also the primary raw material of the various metabolite of microorganism.With carbon source addition it is
0.3% ratio, is that strain growth carries with glucose, sucrose, soluble starch, D-Fructose, beef extract and mannitol respectively
For carbon source, and enter under the conditions of the fixed the most suitable fermentation time of embodiment 2～6, temperature, pH value, inoculum concentration and optimum speed
Row fermentation, detects the impact to Antagonistic Fungi bacteriostatic activity for the different carbon source, determines optimum carbon source.
Experimental results refer to Fig. 6.Can be obtained by Fig. 6, be selected from the carbon source of fermentation medium：Beef extract, mannitol
And during one or more of D-Fructose, the bacteriostasis rate of bacterial strain is all presented on a good bacteriostasis；Wherein, work as fermentation
When the carbon source of culture medium is beef extract, bacteriostatic activity highest.Therefore, the carbon source of optimum fermentation medium is beef extract.
Embodiment 8
The present embodiment is detection in Alternaria alternate Antagonistic Fungi, that is, in the fermentation process of bacillus, fermentation medium
The specific embodiment that carbon source affects for Antagonistic Fungi bacteriostasis rate.
The ratio being 1% with nitrogen source addition, with peptone, tryptone, yeast extract, ammonium oxalate and urea be respectively
Strain growth provides nitrogen source, and in the fixed the most suitable fermentation time of embodiment 2～7, temperature, pH value, inoculum concentration, rotating speed and carbon
Fermented under the conditions of source, detect the impact to Antagonistic Fungi bacteriostatic activity for the different nitrogen sources, determine optimum nitrogen source.
Experimental results refer to Fig. 7.Can be obtained by Fig. 7, be selected from the nitrogen source of fermentation medium：Peptone, yeast leaching
When one or more of cream and tryptone, the bacteriostasis rate of bacterial strain is all presented on a good bacteriostasis；Wherein, when
When the nitrogen source of fermentation medium is peptone, bacteriostatic activity highest.Therefore, the nitrogen source of optimum fermentation medium is albumen
Peptone.
In sum, the invention provides a kind of fermentation process of Alternaria alternate Antagonistic Fungi, described fermentation process is：Institute
State Alternaria alternate Antagonistic Fungi to be inoculated in fermentation medium, shaken cultivation, obtain Alternaria alternate Antagonistic Fungi；Described shaken cultivation
Time be 72h.Present invention also offers a kind of above-mentioned fermentation process cultivates the Alternaria alternate Antagonistic Fungi being obtained resisting in tobacco
The application in sick field.Can obtain after testing, the Alternaria alternate Antagonistic Fungi that the technical scheme cultivation that the present invention provides obtains, bacteriostasis rate
Significantly improve, can be used for the preventing and treating of Alternaria alternate.
The above is only the preferred embodiment of the present invention it is noted that ordinary skill people for the art
For member, under the premise without departing from the principles of the invention, some improvements and modifications can also be made, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (10)
1. a kind of fermentation process of Alternaria alternate Antagonistic Fungi is it is characterised in that described fermentation process is：Described Alternaria alternate
Antagonistic Fungi is inoculated in fermentation medium, shaken cultivation, obtains Alternaria alternate Antagonistic Fungi；
The time of described shaken cultivation is 72h.
2. fermentation process according to claim 1 is it is characterised in that the inoculum concentration of described Alternaria alternate Antagonistic Fungi is
0.1%～2%.
3. fermentation process according to claim 2 is it is characterised in that the inoculum concentration of described Alternaria alternate Antagonistic Fungi is
0.5%.
4. fermentation process according to claim 1 is it is characterised in that the temperature of described shaken cultivation is 25～45 DEG C.
5. fermentation process according to claim 1 is it is characterised in that the initial pH of described fermentation medium is 5～9.
6. fermentation process according to claim 1 is it is characterised in that the rotating speed of described shaken cultivation is 130～170r/
min.
7. fermentation process according to claim 1 is it is characterised in that the carbon source of described fermentation medium is selected from：Beef extract,
One or more of mannitol and D-Fructose.
8. fermentation process according to claim 1 is it is characterised in that the nitrogen source of described fermentation medium is selected from：Peptone,
One or more of yeast extract and tryptone.
9. fermentation process according to claim 1 is it is characterised in that described fermentation process also includes：Separating-purifying；
Described separating-purifying step is carried out after described shaken cultivation.
10. a kind of fermentation process including described in claim 1 to 9 any one is cultivated the Alternaria alternate Antagonistic Fungi being obtained and is sent out
Zymotic fluid is in the application in tobacco disease resistance field.
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| CN201611152844.3A CN106399210A (en) | 2016-12-14 | 2016-12-14 | Fermentation method and application of antagonistic bacteria for tobacco alternaria alternate |
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Citations (3)
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|---|---|---|---|---|
| CN1846505A (en) * | 2006-05-12 | 2006-10-18 | 云南大学 | Tobacco rust resisting microbial prepn and its prepn process and application |
| CN101595893A (en) * | 2009-06-05 | 2009-12-09 | 云南省烟草农业科学研究院 | A kind of bacillus subtilis microbial agent of preventing and treating Alternaria alternate and preparation method thereof |
| CN102154186A (en) * | 2011-04-14 | 2011-08-17 | 中国农业科学院烟草研究所 | Bacillus subtilis and use thereof in prevention and control of fungus disease |
-
2016
- 2016-12-14 CN CN201611152844.3A patent/CN106399210A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1846505A (en) * | 2006-05-12 | 2006-10-18 | 云南大学 | Tobacco rust resisting microbial prepn and its prepn process and application |
| CN101595893A (en) * | 2009-06-05 | 2009-12-09 | 云南省烟草农业科学研究院 | A kind of bacillus subtilis microbial agent of preventing and treating Alternaria alternate and preparation method thereof |
| CN102154186A (en) * | 2011-04-14 | 2011-08-17 | 中国农业科学院烟草研究所 | Bacillus subtilis and use thereof in prevention and control of fungus disease |
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| Title |
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| 关小红 等: "烟草赤星病拮抗细菌Tpb55 摇瓶发酵条件的筛选", 《中国烟草科学》 * |
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Application publication date: 20170215 |