CN104152372B - The biocontrol bacterial strain G68 of controlling plant diseases and its bacterial preparation process and application - Google Patents
The biocontrol bacterial strain G68 of controlling plant diseases and its bacterial preparation process and application Download PDFInfo
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- CN104152372B CN104152372B CN201410247901.0A CN201410247901A CN104152372B CN 104152372 B CN104152372 B CN 104152372B CN 201410247901 A CN201410247901 A CN 201410247901A CN 104152372 B CN104152372 B CN 104152372B
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Abstract
The invention discloses the biocontrol microorganisms and its bacterial preparation process of controlling plant diseases and application.The biocontrol microorganisms are named asBacillus subtilisG68, preserving number CGMCC NO.8978；The preparation method of the biological and ecological methods to prevent plant disease, pests, and erosion bacteria agent is that the bacterial strain G68 after activation is inoculated in 22～26h of NYBE nutrient solution cultures to obtain seed liquor, and seed liquor is inoculated in containing 0.08～0.12 mM MnSO470～74 h of NYBE fermentation mediums culture obtain zymotic fluid, as biological and ecological methods to prevent plant disease, pests, and erosion bacteria agent；Biocontrol agent containing bacterial strain G68 can promote plant seed germination, accelerate plant growth, and to tomato seedling damping-off and early blight diseases control respectively up to 53.8% and 60.1%；Bacterial strain G68 has has a broad antifungal spectrum, and effect stability, preparation cost are low, and application process is simple, environmental friendliness, the features such as nonhazardous, has good application prospect in terms of control of plant disease.
Description
Technical field
The invention belongs to agricultural biological technical field, and in particular to the biocontrol microorganisms of controlling plant diseases and its microbial inoculum preparation side
Method and application.
Background technology
Used as a large agricultural country, the disease control problem of crops is quite important in agricultural planting for China.It is reported that,
China cereal crops are every year because of loss at least 10% ~ 15% that disease problem is caused.The generation of corps diseases often makes country
Economy and people's lives sustain losses severely.The method of control of plant disease has plant quarantine, breeding for disease resistance, cultural control, change
Learn preventing and treating, physics and mechanical control and biological control etc..
Seed selection and using breeding for disease resistance method be controlling disease occur effective way.However, disease-resistant variety is in big face
Product easily in several years lost its disease resistance and becomes infection kind after promoting.Chemical prevention because its produce simplicity, low price,
Particularly instant effect the advantages of, occupy main status in the preventing and treating of plant disease always, and be widely used.But using not
When, then can cause phyto toxicity, murder beneficial microbe, cause pathogen to develop immunity to drugs, the residues of pesticides of high concentration can also
The problems such as poisoning of generation people, animal, environmental pollution.The research of biological control is from the beginning of the thirties in 20th century.Biological control has is permitted
More better than the characteristics of chemical prevention, which will not kill beneficial microbe just for pathogenic microorganism, and to people, animal safety, no
Environmental pollution can be caused.Therefore, biological control is just becoming the important method of control of plant disease, and both at home and abroad biological control is ground
Study carefully more and more, the prevention and controls have broad application prospects.
Bacillus (Bacillus) is a kind of mesophilic, aerobic, sporiferous bacillus, and its physiological characteristic is various, point
Cloth extensively, is easily separately cultured.The bacterium is widespread in nature, nontoxic to people and animals, free from environmental pollution, can produce many
Antibiotic and enzyme are planted, with broad spectrum antibiotic activity and extremely strong anti-adversity ability.Biological prevention and control agent preventing and treating plant is prepared with bacillus
The research of disease becomes the focus of domestic and international research.But the antimicrobial spectrum of different strains, bacteriostasis, antifungal mechanism and antibacterial
Stability of effect etc. also has larger difference.Therefore, it is still biological control to screen the efficient, bacillus of wide spectrum, effect stability
A study hotspot.
The content of the invention
In order to solve the above problems, for using chemical bactericide in a large number in current agricultural production control of plant disease, make
The problems such as residues of pesticides, environmental pollution, the invention provides a kind of efficient, wide spectrum, effect stability and eco-friendly plant
Disease Biocontrol Strain G68.
It is an object of the invention to provide one plant of novel bacterial：Bacillus subtilis（Bacillus subtilis）G68.
Further object is that providing application of the above-mentioned bacterial strains during control of plant disease.
It is yet a further object of the present invention to provide a kind of control of plant disease containing Bacillus subtilis G68
Agent.
It is yet a further object of the present invention to provide the preparation side of biocontrol bacterial strain Bacillus subtilis G68 microbial inoculums
Method.
The technical solution used in the present invention is：
A kind of plant disease biocontrol bacterial strain G68, is identified as bacillus subtilis (Bacillus subtilis), and 2014
On March 28, in is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, and preservation address is court of Beijing
The institute 3 of positive area's North Star West Road 1, Institute of Microorganism, Academia Sinica, deposit number be CGMCC NO.8978, it is proposed that point
Class is named as Bacillus subtilis, has identified on March 28th, 2014 that the bacterial strain of preservation is survival.
Applications of the above-mentioned biocontrol bacterial strain Bacillus subtilis G68 during control of plant disease.
Further, above-mentioned plant disease is vegetative bacteria or fungal disease.
A kind of plant disease controlling agent, which contains above-mentioned plant disease biocontrol bacterial strain Bacillus subtilis G68.
The preparation method of biocontrol bacterial strain Bacillus subtilis G68 microbial inoculums, comprises the following steps：By claim 1
Described biocontrol bacterial strain G68 is inoculated in LB flat boards and activates under the conditions of 29～31 DEG C, and the biocontrol bacterial strain G68 after activation is inoculated in
NYBE nutrient solutions, cultivate 22～26h, obtain seed liquor by 29～31 DEG C under the conditions of 105～115 r/min；Seed liquor is pressed into 2.5
～3.5% inoculum concentration is inoculated in containing 0.08～0.12 mM MnSO4NYBE fermentation mediums, 36～38 DEG C, 190～
70～74 h are cultivated under the conditions of 210r/min, zymotic fluid, i.e. biocontrol bacterial strain Bacillus subtilis G68 microbial inoculums is obtained.
The invention has the beneficial effects as follows：
1）The biocontrol agent preparation process of the present invention is simple, and cost is relatively low, alternative or reduce application to chemical pesticide,
Residues of pesticides are reduced, peasant's agriculture production cost is reduced.The microbial inoculum is biological product, and nonhazardous, non-environmental-pollution are conducive to nothing
The production of public hazards crop and the sustainable development of agricultural.
2）The biocontrol bacterial strain G68 of the present invention shows in laboratory's flat board antagonistic effect：Biocontrol bacterial strain G68 can be vertical to tomato
Rot, early blight of tomato, wheat scab, anthracnose of maize, wax gourd droop, sugarcane black rot, cotton verticillium wilt, Sclerotina Sclerotiorum in Winter Rape
The vegetative bacteria such as the various plants such as core disease fungi and bacterial wilt of tomato pathogen shows good inhibition, with wider
Antimicrobial spectrum.And many Secondary Cultures of Jing, the bacterial strain disease resisting effect are stable.
3）The biocontrol bacterial strain G68 of the present invention shows in greenhouse test result：Life containing biocontrol bacterial strain G68 of the present invention
Anti- microbial inoculum can promote seed to sprout and plant growth, and the biocontrol agent effectively inhibits the growth of pathogen, for tomato
Damping-off（Rhizoctonia solani）And early blight（Alternaria tomatophila）Disease shows significantly
Prevention effect, preventive effect is respectively up to 53.8% and 60.1%.
Description of the drawings
Fig. 1 is that biocontrol bacterial strain G68 suppresses tomato seedling damping-off pathogen（Rhizoctonia solani）Flat board antagonistic effect；
Fig. 2 is that biocontrol bacterial strain G68 suppresses early blight of tomato pathogen（Alternaria tomatophila）Flat board antagonism
Test.
Specific embodiment
With reference to specific embodiment, the present invention is further illustrated, but is not limited thereto.
Embodiment 1
First, the separation of biocontrol bacterial strain G68, screening and identification
A kind of plant disease biocontrol bacterial strain G68, is named as Bacillus subtilis G68, by Guangdong Agricultural science
Institute's agricultural resource is isolated with the compost in Environmental Research Institute's organic solid castoff process base.Which specifically separates, screens,
Authentication method is as follows：
（1）The separation of biocontrol bacterial strain G68
The compost sample that 10.0 g are well mixed is weighed, 90 ml sterilized waters are added, 30 DEG C, 180r/min vibrates 30
Min, bacterium solution is diluted 10 successively-1、10-2、10-3、10-4、10-5、10-6、10-7Times, draw appropriate dilution bacteria suspension（Dilution
10-5、10-6、10-7Bacterium solution again）100 l are uniformly coated on nutrient agar panel, each dilution factor in triplicate, in 30
1 ~ 2d is cultivated in DEG C constant incubator.According to the difference of the physiological characters such as bacterium shape, size, color and luster, picking single bacterium colony is applied again
Flat board further looks at its form until the bacterial strain of the single form of acquisition finally by microscopy, really as pure for single form
Change, 4 DEG C of refrigerations are standby after purification.
（2）The screening of biocontrol bacterial strain G68
The screening of biocontrol bacterial strain is carried out using flat board antagonistic effect method, it is 1 × 10 to draw concentration5The pathogen of CFU/ml
100 l of bacterium solution is respectively put into LB culture mediums（Bacterium）Or PDA culture medium（Fungi）It is coated with uniformly, using aseptic punching in flat board
Device makes a call to 4 holes on flat board, and nutrient solution, the fermentation supernatant of the different isolated strains that 24h are cultivated at 30 DEG C are injected separately in every hole
Liquid（6000 r/min centrifugation 30min remove thalline）, sterilized water and 1mM 80 l of cycloheximide, each process is in triplicate.
After 3 ~ 5 d are cultivated in 30 DEG C of constant incubators, observation whether there is inhibition zone, calculates bacteriostatic activity.
Bacteriostatic activity is represented with antibacterial circle diameter:Antibacterial circle diameter=transparent loop diameter-aperture (8 mm).
By the bacterial strain with bacteriostatic activity, on LB flat boards, 10 times () is cultivated in continuous switching every 7d once, is determined and is passed every time
The antagonistic stability of culture bacterial strain.
According to phytopathogen tomato seedling damping-off, early blight of tomato, bacterial wilt of tomato, wheat scab, corn anthrax
The bacteriostatic activity of disease, wax gourd droop, sugarcane black rot, cotton verticillium wilt, sclerotinia rot of colza etc. and pass on rear bacteriostatic activity
Stability, filter out the bacterial strain of a plant height effect, wide spectrum, the plant epiphyte resisting of effect stability and bacterial disease opportunistic pathogen, bacterial strain is pure
After change, ~ 70 DEG C of ultra low temperature freezers are stored in 50% glycerine.
The identification of biocontrol bacterial strain G68
Jing forms, dyeing, biochemical reactions and molecular biology identification are Bacillus subtilis.
A. the colonial morphology of biocontrol bacterial strain G68 and thalline feature
On LB solid mediums, colonial morphology is less, edge is irregular, surface is moistened without projection, relatively, opaque, in breast
White.Somatic cells be in rod-short, even dyeing, gram-positive bacteria, central spore, peritrichous.
B. the physiological and biochemical property of biocontrol bacterial strain G68 of the present invention
The physiological and biochemical property of biocontrol bacterial strain G68 is：Aerobism, glucose fermentation type, catalase, Starch Hydrolysis, nitric acid
Salt reduction, casein hydrolysis, gelatin hydrolysis, glycitols fermentation, M.R. and V-P result of the tests are the positive；Citrate hydrolysis, fiber
Element decomposes and tyrosine hydrolysis test result is feminine gender（As shown in table 1）.
The physiological and biochemical property of 1 biocontrol bacterial strain G68 of table
Note："+" is positive, and "-" is negative.
C. the molecular biology identification of biocontrol bacterial strain G68
Bacterial genomes DNA are extracted using Fast DNA kit, is sequenced using the universal primer of 16srRNA.It is used
Primer sequence is：8F (5'-AGAGTTTGA TCATGG CTCAG-3') and 907R (5'-CCGTCAATTCCTTTGAGTTT-
16S rDNA genetic fragments are amplified from genomic DNA 3').The sequencing of PCR primer commission TaKaRa companies, sequencing result are submitted to
Accession number KJ123643 is obtained to GenBank databases, using Blast program search homologous sequences, as a result with accession number
NR104873 Bacillus subtilis subsp. inaquosorum strain BGSC 3A28 homologys are 99%, because
This, with reference to colonial morphology and physiological and biochemical property, biocontrol bacterial strain G68 is accredited as Bacillus subtilis.
2nd, the preparation of biocontrol bacterial strain G68 microbial inoculums
The biocontrol bacterial strain G68 of LB slant mediums will be stored in, the activation of LB flat boards will be inoculated in（30℃）, by the life after activation
Anti- bacterial strain G68 is inoculated in NYBE（8 g nutrient broths, 7 g dusty yeasts, 1L H2O）Nutrient solution, 30 DEG C, under the conditions of 110 r/min
Culture 24h, obtains seed liquor.Seed liquor is inoculated in into NYBE.Mn by 3% inoculum concentration（NBYE+0.1 mM MnSO4·H2O）Send out
Ferment culture medium（8 g nutrient broths, 7 g dusty yeasts, 0.1mM MnSO4·H2O, 1L H2O）, 37 DEG C, train under the conditions of 200 r/min
72 h are supported, zymotic fluid is obtained（Biological and ecological methods to prevent plant disease, pests, and erosion bacterium solution）, i.e. biocontrol bacterial strain G68 microbial inoculums, wherein biocontrol bacterial strain G68 viable counts are 109~
1010CFU/ml。
3rd, inhibitory action of the biocontrol bacterial strain G68 to phytopathogen
（1）Flat board antagonistic effect
It is 1 × 10 to draw concentration5The phytopathogen of CFU/ml（As shown in table 2）100 l of bacterium solution is respectively put into LB（Carefully
Bacterium）Or PDA（Fungi）It is coated with uniformly in flat board, 4 holes is made a call on each flat board using aseptic card punch, 80 l are injected separately into
Sterilized water, the cycloheximide of 1mM, the fermented supernatant fluid of biocontrol bacterial strain G68 microbial inoculums and biocontrol bacterial strain G68 microbial inoculums, each process
In triplicate, after cultivating 3～5 days in 30 DEG C of constant incubators, observation whether there is inhibition zone, and determines inhibition zone activity, antibacterial work
Property antibacterial circle diameter（mm）Represent：Antibacterial circle diameter=transparent loop diameter-aperture (8 mm).
The measurement result of inhibition zone activity is as shown in table 2 below, it can be seen that bacterial strain G68 of the present invention and its fermentation supernatant
Liquid is to common phytopathogen tomato seedling damping-off bacterium, tomato early blight bacterium, fusarium graminearum, anthracnose of maize bacterium, wax gourd
Wilt, sugarcane black rot, sugarcane black rot, verticillium dahliae, Sclerotinia sclerotiorum and bacterial wilt of tomato bacterium are equal
With obvious fungistatic effect, its fungistatic effect is substantially better than the fungistatic effect of cycloheximide.Illustrate the biocontrol bacterial strain of the present invention
G68 shows good inhibition to various plants fungi and bacterial disease opportunistic pathogen, with wider antimicrobial spectrum.
Wherein bacterial strain G68 and its fermented supernatant fluid are to tomato seedling damping-off pathogen, the antibacterial figure of early blight of tomato pathogen
Respectively as depicted in figs. 1 and 2, it can be seen that bacterial strain G68 and its fermented supernatant fluid are generated to both phytopathogens
Significantly inhibition zone, the especially inhibition zone of bacterial strain G68 are significantly greater than the inhibition zone of cycloheximide.
In sum, biocontrol bacterial strain G68 of the invention has inhibitory action very to frequently seen plants pathogen.
Antibacterial circle diameters of the 2 biocontrol bacterial strain G68 of table to phytopathogen（mm）
（2）Greenhouse test determines application effect of the biocontrol bacterial strain G68 microbial inoculums in anti-tomato seedling damping-off
The biocontrol bacterial strain G68 microbial inoculums prepared in taking above-mentioned " two ", biocontrol bacterial strain G68 viable counts are 1 × 109CFU/ml；Kind
Eggplant damping-off pathogen is inoculated in PDA culture medium, cultivates 7d, and with aseptic water washing, it is 1 × 10 to adjust spore concentration5CFU/ml。
5 process are set altogether：
Treatment group 1：24 h of tomato seeds is soaked with biocontrol bacterial strain G68 microbial inoculums, be after planting inoculated with 5ml tomato seedling damping-off cause of diseases
Bacterium and 5ml sterilized waters；
Treatment group 2：Seed is after planting inoculated with 5ml biocontrol bacterial strain G68 microbial inoculums and tomato seedling damping-off pathogen respectively；
Treatment group 3（Control group）：Seed is after planting inoculated with 5ml tomato seedling damping-offs pathogen and 5ml sterilized waters；
Treatment group 4：Seed is after planting inoculated with 5ml biocontrol bacterial strain G68 microbial inoculums and 5ml sterilized waters；
Treatment group 5（Conventional group）：Seed is after planting inoculated with 10ml sterilized waters.
150 g soil are housed per basin, 10 seeds are sowed；Each processes 6 repetitions, and above-mentioned tomato seeds are seeded in soil
In earth, tomato is positioned over into temperature for 28 DEG C, illumination is culture under the conditions of 16h/8h, and observation germination situation and disease are anti-
Effect.And count the incidence of disease, and onset grade is divided into into 0,1,2,3 totally four grades：
0 grade：Do not fall ill；
1 grade：Seedling basal part of stem produces oval crineous scab；
2 grades：Scab is gradually recessed, around stem 1 week after expansion；
3 grades：Basal part of stem is in ulcer shape, is wilted, so that it is dead；
Disease index=［ ∑（Diseased plant numbers at different levels × corresponding onset grade value）/ investigation total strain number × highest onset grade value ］
×100%；
Preventive effect=（The disease index of control group disease index-treatment group）/ control group disease index ］ × 100%.
3 each disease index for processing of table and preventive effect feelings
As a result show, 4 seed for the treatment of group is sprouted earlier than other treatment groups for one day, and growth of seedling speed is fast.Treatment group 1 and place
Reason group 2 serves preferable disease control effect, and the prevention effect of two kinds of processing methods is respectively 51.3% and 53.8%, has no
Significant difference.
Above-mentioned result of the test explanation, biocontrol bacterial strain G68 of the present invention have good disease control effect to plant.
（3）Greenhouse test determines application effect of the biocontrol bacterial strain G68 microbial inoculums in anti-early blight of tomato
The biocontrol bacterial strain G68 microbial inoculums prepared in taking above-mentioned " two ", biocontrol bacterial strain G68 viable counts are 1 × 1010CFU/ml；Kind
Eggplant early blight pathogen is inoculated in PDA culture medium, cultivates 7 d, with aseptic water washing, adjusts spore concentration 1 × 106CFU/
ml.Tomato first carries out nursery, and band grew to for 4 leaf phases and transplanted.4 process are set：
Treatment group 1：50ml biocontrol agents will be poured after seedling replanting, after 3 days, pour 30ml early blight of tomato pathogens；
Treatment group 2（Control group）：50ml sterilized waters are poured after transplanting, 30ml early blight of tomato pathogens after 3d, are poured；
Treatment group 3：50ml biocontrol agents will be poured after seedling replanting, after 3d, pour 30ml sterilized waters；
Treatment group 4（Conventional group）：50ml sterilized waters are poured after transplanting, 30ml sterilized waters after 3d, are poured again.
Per 3 seeds of basin, each processes 5 repetitions, and tomato is positioned over temperature for 28 DEG C, and illumination is under the conditions of 16h/8h
Culture, observation tomato plant strain growth situation and diseases control.And count the incidence of disease：Be divided into 0,1,2,3,4,5,6 totally seven send out
Sick grade：
0 grade：Do not fall ill；
1 grade：1 blade morbidity, has fragmentary scab on sick leaf, scab accounts for less than the 10% of blade face area；
2 grades：2 ~ 4 blade morbidities, have a small amount of scab on sick leaf, scab accounts for less than the 11% ~ 25% of blade face area；
3 grades：5 ~ 7 blade morbidities, on sick leaf, Lesion number is medium, and scab accounts for less than the 26% ~ 50% of blade face area；
4 grades：A large amount of blade morbidities, on sick leaf, Lesion number is more, and scab accounts for less than the 51% ~ 75% of blade face area；
5 grades：A large amount of blade morbidities, whole blade are withered；
6 grades：Plant is dead.
Disease index=［ ∑（Diseased plant numbers at different levels × corresponding onset grade value）/ investigation total strain number × highest onset grade value ］
×100%；
Preventive effect=（The disease index of control group disease index-treatment group）/ control group disease index ］ × 100%.
As a result show：Biocontrol bacterial strain G68 has growth-promoting functions to tomato, transplants 25 d and post-processes 3 tomato growth plant relatively place
4 high 10cm are managed, stem slightly relatively processes 4 width 0.2mm.Meanwhile, biocontrol bacterial strain G68 microbial inoculums show preferably preventing and treating to early blight of tomato
Effect, preventive effect is up to 60.1%.
The disease index and preventive effect situation of 4 each treatment group of table
Above-mentioned result of the test explanation, biocontrol bacterial strain G68 of the present invention have good disease control effect to plant.
Claims (4)
1. the biocontrol bacterial strain bacillus subtilis of a kind of controlling plant diseases（Bacillus subtilis）G68, is deposited in China
General Microbiological Culture preservation administrative center, its preserving number are CGMCC NO.8978.
2. applications of the biocontrol bacterial strain bacillus subtilis G68 described in claim 1 during control of plant disease；
Described plant disease is vegetative bacteria or fungal disease；
The fungal diseases of plants is tomato seedling damping-off bacterium, tomato early blight bacterium, fusarium graminearum, anthracnose of maize bacterium, winter
Cucurbit wilt bacterium, sugarcane black rot, verticillium dahliae, Sclerotinia sclerotiorum, the phytobacterial disease are that tomato green grass or young crops is withered
Germ.
3. a kind of plant disease controlling agent, it is characterised in that：It is withered containing the biocontrol bacterial strain described in claim 1 in the control agent
Careless bacillus G68.
4. the preparation method of the microbial inoculum of the biocontrol bacterial strain bacillus subtilis G68 described in claim 1, it is characterised in that：Including
Following steps：Biocontrol bacterial strain bacillus subtilis G68 described in claim 1 is inoculated in into LB flat boards under the conditions of 29～31 DEG C
Biocontrol bacterial strain G68 after activation is inoculated in NYBE nutrient solutions, 29～31 DEG C, is cultivated under the conditions of 105～115 r/min by activation
22～26h, obtains seed liquor；Seed liquor is inoculated in containing 0.08～0.12 mM MnSO by 2.5～3.5% inoculum concentration4's
NYBE fermentation mediums, cultivate 70～74 h, obtain zymotic fluid, i.e. biocontrol bacterial strain by 36～38 DEG C under the conditions of 190～210r/min
The microbial inoculum of bacillus subtilis G68；
Described NYBE nutrient solutions are by 8g nutrient broths, 7g dusty yeasts and 1L H2O is constituted；
The NYBE fermentation mediums are by 8g nutrient broths, 7g dusty yeasts, 0.08～0.12 mM MnSO4With 1L H2O is constituted.
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| CN105543121A (en) * | 2015-11-17 | 2016-05-04 | 河南科技学院 | Bacillus subtilis and application thereof in maize anthracnose bio-control |
| CN106434462B (en) * | 2016-10-12 | 2019-05-10 | 山东省林业科学研究院 | One bacillus subtilis and its application in terms of prevention and treatment blueberry anthracnose, diseases of mulberry fruits |
| CN108651525A (en) * | 2018-07-16 | 2018-10-16 | 广西南宁益土生物科技有限责任公司 | A kind of granular pesticide containing bacillus subtilis and humic acid |
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| CN102925379A (en) * | 2012-08-02 | 2013-02-13 | 中南林业科技大学 | Bacillus licheniformis used for controlling cunninghamia lanceolata anthracnose, and application thereof |
| CN103173394A (en) * | 2013-03-21 | 2013-06-26 | 西南大学 | Bacillus subtilis for controlling citrus canker |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102925379A (en) * | 2012-08-02 | 2013-02-13 | 中南林业科技大学 | Bacillus licheniformis used for controlling cunninghamia lanceolata anthracnose, and application thereof |
| CN103173394A (en) * | 2013-03-21 | 2013-06-26 | 西南大学 | Bacillus subtilis for controlling citrus canker |
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| GenBank Accession：KJ123643.1;Gu W;《GenBank》;20140329;第1页 * |
| 地衣芽孢杆菌生防菌株SDYT-79发酵条件优化;纪明山等;《沈阳农业大学学报》;20110430;第42卷(第2期);第164-169页 * |
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