CN105907683A - Bacillus amyloliquefaciens cd5bc and application thereof - Google Patents
Bacillus amyloliquefaciens cd5bc and application thereof Download PDFInfo
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- CN105907683A CN105907683A CN201610390489.7A CN201610390489A CN105907683A CN 105907683 A CN105907683 A CN 105907683A CN 201610390489 A CN201610390489 A CN 201610390489A CN 105907683 A CN105907683 A CN 105907683A
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- 241000193744 Bacillus amyloliquefaciens Species 0.000 title claims abstract description 28
- 201000010099 disease Diseases 0.000 claims abstract description 34
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 34
- 240000008067 Cucumis sativus Species 0.000 claims abstract description 11
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 claims abstract description 11
- 240000007594 Oryza sativa Species 0.000 claims abstract description 11
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 11
- 235000009566 rice Nutrition 0.000 claims abstract description 11
- 230000003405 preventing effect Effects 0.000 claims abstract description 9
- 235000009811 Momordica charantia Nutrition 0.000 claims abstract description 6
- 244000078912 Trichosanthes cucumerina Species 0.000 claims abstract description 6
- 235000008322 Trichosanthes cucumerina Nutrition 0.000 claims abstract description 6
- 206010039509 Scab Diseases 0.000 claims abstract description 5
- 238000004321 preservation Methods 0.000 claims abstract description 4
- 238000002360 preparation method Methods 0.000 claims abstract description 3
- 241000607479 Yersinia pestis Species 0.000 claims description 15
- 238000000034 method Methods 0.000 claims description 15
- 235000002566 Capsicum Nutrition 0.000 claims description 12
- 240000008574 Capsicum frutescens Species 0.000 claims description 12
- 239000001390 capsicum minimum Substances 0.000 claims description 12
- 241000196324 Embryophyta Species 0.000 claims description 11
- 239000007788 liquid Substances 0.000 claims description 11
- 238000000855 fermentation Methods 0.000 claims description 10
- 230000004151 fermentation Effects 0.000 claims description 10
- 210000000582 semen Anatomy 0.000 claims description 10
- UHPMCKVQTMMPCG-UHFFFAOYSA-N 5,8-dihydroxy-2-methoxy-6-methyl-7-(2-oxopropyl)naphthalene-1,4-dione Chemical compound CC1=C(CC(C)=O)C(O)=C2C(=O)C(OC)=CC(=O)C2=C1O UHPMCKVQTMMPCG-UHFFFAOYSA-N 0.000 claims description 9
- 241000894006 Bacteria Species 0.000 claims description 9
- 241000223218 Fusarium Species 0.000 claims description 9
- 239000003814 drug Substances 0.000 claims description 8
- 230000003628 erosive effect Effects 0.000 claims description 8
- 206010025421 Macule Diseases 0.000 claims description 5
- 230000000844 anti-bacterial effect Effects 0.000 claims description 5
- 239000002689 soil Substances 0.000 claims description 4
- 241000272525 Anas platyrhynchos Species 0.000 claims description 3
- 241000193830 Bacillus <bacterium> Species 0.000 claims description 2
- 239000012531 culture fluid Substances 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 241001530056 Athelia rolfsii Species 0.000 claims 1
- 229920002472 Starch Polymers 0.000 claims 1
- 235000019698 starch Nutrition 0.000 claims 1
- 239000008107 starch Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 28
- 230000000443 biocontrol Effects 0.000 abstract description 21
- 238000011081 inoculation Methods 0.000 abstract description 8
- 240000006677 Vicia faba Species 0.000 abstract description 5
- 235000010749 Vicia faba Nutrition 0.000 abstract description 5
- 235000002098 Vicia faba var. major Nutrition 0.000 abstract description 5
- 244000052616 bacterial pathogen Species 0.000 abstract description 3
- 240000004160 Capsicum annuum Species 0.000 abstract 2
- 235000008534 Capsicum annuum var annuum Nutrition 0.000 abstract 2
- 235000007862 Capsicum baccatum Nutrition 0.000 abstract 2
- 239000001728 capsicum frutescens Substances 0.000 abstract 2
- 235000019219 chocolate Nutrition 0.000 abstract 2
- 230000001276 controlling effect Effects 0.000 abstract 2
- 241000233614 Phytophthora Species 0.000 abstract 1
- 241000221662 Sclerotinia Species 0.000 abstract 1
- 230000002265 prevention Effects 0.000 description 11
- 230000001580 bacterial effect Effects 0.000 description 9
- 244000005700 microbiome Species 0.000 description 7
- QNBTYORWCCMPQP-JXAWBTAJSA-N (Z)-dimethomorph Chemical compound C1=C(OC)C(OC)=CC=C1C(\C=1C=CC(Cl)=CC=1)=C/C(=O)N1CCOCC1 QNBTYORWCCMPQP-JXAWBTAJSA-N 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 3
- 239000005761 Dimethomorph Substances 0.000 description 3
- 241000233616 Phytophthora capsici Species 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 230000000813 microbial effect Effects 0.000 description 3
- 244000052769 pathogen Species 0.000 description 3
- 230000001717 pathogenic effect Effects 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000001408 fungistatic effect Effects 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 238000007689 inspection Methods 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- FRXSZNDVFUDTIR-UHFFFAOYSA-N 6-methoxy-1,2,3,4-tetrahydroquinoline Chemical compound N1CCCC2=CC(OC)=CC=C21 FRXSZNDVFUDTIR-UHFFFAOYSA-N 0.000 description 1
- 241000194110 Bacillus sp. (in: Bacteria) Species 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 208000035199 Tetraploidy Diseases 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 244000000037 crop pathogen Species 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000706 filtrate Substances 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 230000001788 irregular Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000003900 soil pollution Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000012549 training Methods 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/07—Bacillus
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- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/10—Animals; Substances produced thereby or obtained therefrom
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Abstract
The invention provides bacillus amyloliquefaciens cd5bc. The strain is classified and named as Bacillus amyloliquefaciens cd5bc and is preserved in China Center for Type Culture Collection, the preservation number is CCTCC NO:M 2016232, and the preservation time is April 28<th>, 2016. The invention further provides application of the strain in the aspect of preventing and controlling crop diseases and a biocontrol preparation prepared from the strain. The strain has a very significant preventing and controlling effect on red pepper phytophthora blight, the biocontrol effect reaches up to 100% after inoculation is conducted for 15 days, and the biocontrol effect reaches up to 82.36% after inoculation is conducted for 60 days. The bacillus amyloliquefaciens cd5bc has excellent disease-resistant effects on multiple crop diseases, the bacteriostasis rates of the strain on pathogenic bacteria of red pepper sclerotinia blight, bitter gourd wilt, cucumber wilt, cucumber scabs, rice blast, a rice sheath blight disease, broad bean chocolate spots and broad bean brown spots all reach up to 70% or above, and the bacteriostasis rate of the strain on the broad bean chocolate spots and broad bean brown spots reaches up to 100%.
Description
Technical field
The invention belongs to field of biological control, relate to bacillus amyloliquefaciens cd5bc and an application thereof, be specifically related to a strain, for capsicum epidemic disease, there is notable prevention effect and the bacillus amyloliquefaciens also various crop disease with efficient prevention effect simultaneously, the biological and ecological methods to prevent plant disease, pests, and erosion medicament obtained based on it and application thereof.
Background technology
In the last few years, utilize the drawback of Agro-chemicals control crop pest gradually to display, mainly show as damaging soil property, causing soil pollution and pesticide residues etc..In this context, biological prevention, in particular with microbial control crop pest technology, increasingly come into one's own.
But, the microorganism being currently used for preventing and treating crop pest is faced with the situation that quantity is very few, effect is relatively low, limits the application of its reality.
Capsicum epidemic disease, Fructus Capsici sclerotiniose, bitter gourd wilt, cucumber fusarium axysporum, yellow scab, rice blast, rice sheath blight disease, Semen Viciae fabae reddish macules and Semen Viciae fabae brown spot make common crop pest, the production to agricultural all the time cause large effect.
In prior art, although sporadicly there being the report of the bacterial strain that can prevent and treat certain crop pest, such as " interior raw bacillus amyloliquefaciens TB2 strain activity material induction Fructus Capsici fruit antilemic Biochemical Mechanism ", but its prevention effect is the most unsatisfactory, and also needs to carry out the extraction operation of complicated active substance.In terms of research can efficiently prevent and treat various crop disease bacterial strain simultaneously, prior art the most not yet obtains gratifying effect, is mainly manifested in the prevention effect difference to various crops excessive, is unfavorable for promoting the use of.
Summary of the invention
Shortcoming for prior art, an object of the present invention is to provide a bacillus amyloliquefaciens cd5bc, the Classification And Nomenclature of this bacillus amyloliquefaciens cd5bc is Bacillus amyloliquefaciens cd5bc, it is preserved in China typical culture collection center (Luo Jia Shan, wuchang, wuhan China typical culture collection center (Wuhan University), postcode: 430072), preserving number is CCTCC NO:M 2016232, and the preservation time is on April 28th, 2016.
The present inventor's long campaigns utilizes the work in terms of microbial control crop pest, grope through substantial amounts of experiment, and inventor is isolated bacillus amyloliquefaciens cd5bc from Broad Bean Leaves.As shown in the embodiment of the present invention, the bacillus amyloliquefaciens cd5bc of the present invention all has efficient fungistatic effect and biocontrol effect for various crop pathogen.
Skilled addressee readily understands that, the present invention is to have found a kind of prevention effect to capsicum epidemic disease and is up to more than 80% (being better than chemical pesticide 50% dimethomorph WG) one of prior art contribution, and multiple to other (at least 8 kinds) crop pest can also have the microbial strains of efficient prevention effect.
Further object is that the application providing above-mentioned bacillus amyloliquefaciens cd5bc in preventing and treating capsicum epidemic disease, Fructus Capsici sclerotiniose, bitter gourd wilt, cucumber fusarium axysporum, yellow scab, rice blast, rice sheath blight disease, Semen Viciae fabae reddish macules, Semen Viciae fabae brown spot on any one.
Further object is that and the biological and ecological methods to prevent plant disease, pests, and erosion medicament prepared by above-mentioned bacillus amyloliquefaciens cd5bc is provided.Described biological and ecological methods to prevent plant disease, pests, and erosion medicament includes the one in ferment product, antibacterial freeze dried powder.
In described ferment product, the cell concentration of antibacterial is 109Cfu/mL～1010cfu/mL。
The preparation method of described ferment product is: after activation biocontrol bacteria cd5bc, dips bacterium solution with the sterilizing rifle head of 200 μ L, then connects first of rifle as shaking training, 150r/min, shaken cultivation 24h at 30 DEG C in NB culture fluid, obtain the fermentation liquid of biocontrol microorganisms.
When preventing and treating capsicum epidemic disease, the method for described application comprises the steps:
When Hot Pepper Seedling grows 4 leaf, prick a duck eye at the soil away from rhizosphere 1～2cm, the fermentation liquid of bacillus amyloliquefaciens cd5bc is inoculated and flows into Fructus Capsici root, every strain 5mL.
When preventing and treating cucumber fusarium axysporum, the method for described application comprises the steps:
When Fructus Cucumidis sativi grows two panels true leaf, with the fermentation liquid pouring root of bacillus amyloliquefaciens cd5bc, every strain 25mL.
Preferably, in above-mentioned application process, in the fermentation liquid of described bacillus amyloliquefaciens cd5bc, cell concentration is 109Cfu/mL～1010cfu/mL。
Beneficial effects of the present invention:
1, bacterial strain of the present invention has the most significant prevention effect for capsicum epidemic disease, and after inoculating 15 days, biocontrol effect is up to 100%, and after inoculating 60 days, biocontrol effect is up to 82.36%；
2, the present invention all has outstanding disease resistance effect to various crop disease, to Fructus Capsici sclerotiniose, bitter gourd wilt, cucumber fusarium axysporum, yellow scab, rice blast, rice sheath blight disease, Semen Viciae fabae reddish macules, the bacteriostasis rate all up to more than 70% of pathogen of Semen Viciae fabae brown spot, the bacteriostasis rate of Semen Viciae fabae reddish macules, Semen Viciae fabae Pathogenic Bacteria Causing Brown Blotch Disease is up to 100%.
Accompanying drawing explanation
Fig. 1 is the cultural colony figure of bacterial strain of the present invention；
Fig. 2 is the sediments microscope inspection result figure of bacterial strain of the present invention；
Fig. 3 is present system developmental analysis figure.
Detailed description of the invention
Below by embodiment, the present invention is specifically described; be necessary it is pointed out here that be that following example are only intended to be further detailed the present invention; it is not intended that limiting the scope of the invention; some nonessential improvement and adjustment that the person skilled in the art in this field is made according to foregoing invention content, still fall within protection scope of the present invention.
Embodiment 1
The form of strains B. amyloliquefaciens cd5bc of the present invention (under be called for short cd5bc), cultural characteristic:
Cd5bc is in faint yellow bacterium colony on NA base, and bacterium colony is flat, circular or subcircular, infiltrates band gloss, and edge is irregular.Gram’s staining is positive, shaft-like, the blunt circle in two ends, has spore, sporangium not to expand, and spore is oval (Fig. 1).Tentatively judge that according to morphological characteristic bacterium cd5bc belongs to bacillus (Bacillus sp.).Sediments microscope inspection, such as Fig. 2.
Embodiment 2
16S rDNA sequence analysis:
After order-checking splicing, the 16S a length of 1466bp of rDNA sequence of bacterial strain cd5bc amplification, by the BLASTN in NCBI website, the 16SrDNA sequence of bacterial strain cd5bc is carried out tetraploid rice, and download sequence with the 16S rDNA sequence of known bacterial strain in gene bank, utilize MEGA to set up phylogenetic tree.Result shows nearest with the sibship of biocontrol microorganisms cd5bc and Bacillus amyloliquefaciens.According to morphological characteristic and 16S rDNA sequential system developmental analysis result, it is determined that cd5bc is bacillus amyloliquefaciens Bacillus amyloliquefaciens.
Embodiment 3
Cd5bc biocontrol effect detects
1. flat board fungistatic effect
It is connected on PDA plate cultivate 5 days by pathogen bacterium phytophthora blight of pepper, Fructus Capsici hyphal cluster germ, bitter gourd wilt, bacterium cake is bought from colony edge with the card punch of a diameter of 5mm, it is placed in blank PDA plate central authorities, the fermentation liquid (line) of biocontrol bacterial strain is accessed on equidistant point of both sides, 26.5 DEG C of constant temperature culture, when pathogen bacterium mycelia will cover with full ware in comparison ware, measure the growth radius of thalline, and calculating suppression ratio, every strain sets 4 repetitions for examination bacterium.Bacteriostasis rate is calculated according to inhibition zone size.
The results are shown in Table 1.
The table 1cd5bc biocontrol effect to various pathogenic bacteria
The 2.cd5bc potted plant biocontrol effect to capsicum epidemic disease
When Hot Pepper Seedling grows 4 leaf, the fermentation liquid of biocontrol microorganisms cd5bc is inoculated in Fructus Capsici root；Every strain 5mL, pricks a duck eye at the soil away from rhizosphere 1～2cm, is injected by fermentation liquid；With 1000 times of liquid of chemical pesticide 50% dimethomorph WG as parallel control, and blank is set；Each process inoculates 30 strains, repeats for 3 times.
After biocontrol microorganisms inoculation 2d, at the position inoculation phytophthora blight of pepper that inoculation biocontrol microorganisms is identical, inoculum density is 104Zoospore/mL, inoculum concentration is 3mL.After inoculation phytophthora blight of pepper 15,30d and 60d, the disease index of investigation capsicum epidemic disease, and calculate the biocontrol effect of biocontrol microorganisms.
Result shows: after inoculation 60d, cd5bc reaches 82.36% to the prevention effect of capsicum epidemic disease, is significantly higher than the prevention effect of 50% dimethomorph WG.
The table 2cd5bc potted plant biocontrol effect to capsicum epidemic disease
The 3.cd5bc potted plant biocontrol effect to cucumber fusarium axysporum
When Fructus Cucumidis sativi grows two panels true leaf, by ferment filtrate pouring root 25mL of cd5bc；Root incised wound inocalation method inoculation cucumber fusarium axysporum pathogen is used, 7,14 and 28d investigation incidence after perfusion, and investigation prevention effect after 24h.Result is as shown in table 3.
The table 3cd5bc biocontrol effect to cucumber fusarium axysporum
Claims (9)
1. a bacillus amyloliquefaciens cd5bc, it is characterised in that described bacillus amyloliquefaciens
The Classification And Nomenclature of cd5bc is Bacillus amyloliquefaciens cd5bc, is preserved in Chinese Typical Representative
Culture collection center, preserving number is CCTCC NO:M 2016232, and the preservation time is 2016
On April 28, in.
2. bacillus amyloliquefaciens cd5bc described in claim 1 is preventing and treating capsicum epidemic disease, Fructus Capsici bacterium
Core disease, bitter gourd wilt, cucumber fusarium axysporum, yellow scab, rice blast, rice banded sclerotial blight
Application on any one in disease, Semen Viciae fabae reddish macules, Semen Viciae fabae brown spot.
3. the biological and ecological methods to prevent plant disease, pests, and erosion medicament prepared by bacillus amyloliquefaciens cd5bc described in claim 1.
Biological and ecological methods to prevent plant disease, pests, and erosion medicament the most according to claim 3, it is characterised in that described biological and ecological methods to prevent plant disease, pests, and erosion medicament bag
Include the one in ferment product, antibacterial freeze dried powder.
Biological and ecological methods to prevent plant disease, pests, and erosion medicament the most according to claim 4, it is characterised in that described ferment product
In, the cell concentration of antibacterial is 109Cfu/mL～1010cfu/mL。
Biological and ecological methods to prevent plant disease, pests, and erosion medicament the most according to claim 4, it is characterised in that described ferment product
Preparation method be: by described antibacterial activate after, inoculate in NB culture fluid, in 150r/min,
Shaken cultivation 24h at 30 DEG C, obtains described ferment product.
Application the most according to claim 2, it is characterised in that when preventing and treating capsicum epidemic disease,
The method of described application comprises the steps:
When Hot Pepper Seedling grows 4 leaf, prick a duck eye at the soil away from rhizosphere 1～2cm,
The fermentation liquid of bacillus amyloliquefaciens cd5bc is inoculated and flows into Fructus Capsici root, every strain 5mL.
Application the most according to claim 2, it is characterised in that when preventing and treating cucumber fusarium axysporum,
The method of described application comprises the steps:
When Fructus Cucumidis sativi grows two panels true leaf, by the fermentation liquid pouring root of bacillus amyloliquefaciens cd5bc,
Every strain 25mL.
9. according to the application described in claim 7 or 8, it is characterised in that described solution starch spore
In the fermentation liquid of bacillus cd5bc, cell concentration is 109Cfu/mL～1010cfu/mL。
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520566A (en) * | 2016-10-27 | 2017-03-22 | 山东农业大学 | Antagonistic fungus for preventing and treating bitter gourd wilt and application thereof |
| CN107217016A (en) * | 2017-05-04 | 2017-09-29 | 华南农业大学 | One plant has the endophytic Bacillus bacterial strain ZY122 for suppressing rice sheath blight disease and its application |
| CN110521738A (en) * | 2019-07-19 | 2019-12-03 | 湖南科技学院 | A kind of compound biological prevention and control agent and its control method for preventing and treating Phytophthora capsici |
| CN111713206A (en) * | 2020-06-19 | 2020-09-29 | 四川省农业科学院植物保护研究所 | Method for increasing yield of bitter gourds by using biocontrol strain bacillus amyloliquefaciens Bam22 |
| CN113151075A (en) * | 2021-04-09 | 2021-07-23 | 安徽农业大学 | Bacillus amyloliquefaciens OR2-30 strain and application thereof |
| CN114365749A (en) * | 2022-01-27 | 2022-04-19 | 云南农业大学 | Composition for preventing and treating black rot and black shank of tobacco root and improving character and quality |
| CN114395397A (en) * | 2022-01-27 | 2022-04-26 | 云南农业大学 | Microbial soil improvement microbial inoculum for preventing and treating tobacco blight and preparation method thereof |
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