CN111748498A - Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof - Google Patents
Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof Download PDFInfo
- Publication number
- CN111748498A CN111748498A CN202010652997.4A CN202010652997A CN111748498A CN 111748498 A CN111748498 A CN 111748498A CN 202010652997 A CN202010652997 A CN 202010652997A CN 111748498 A CN111748498 A CN 111748498A
- Authority
- CN
- China
- Prior art keywords
- bacillus
- parts
- beef extract
- culture
- bacillus amyloliquefaciens
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 241000193830 Bacillus <bacterium> Species 0.000 title claims abstract description 76
- 150000001875 compounds Chemical class 0.000 title claims abstract description 50
- 239000002068 microbial inoculum Substances 0.000 title claims abstract description 50
- 241000221785 Erysiphales Species 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title claims abstract description 23
- 244000005700 microbiome Species 0.000 claims abstract description 3
- 241000193744 Bacillus amyloliquefaciens Species 0.000 claims description 55
- 235000015278 beef Nutrition 0.000 claims description 47
- 239000001888 Peptone Substances 0.000 claims description 42
- 108010080698 Peptones Proteins 0.000 claims description 42
- 235000019319 peptone Nutrition 0.000 claims description 42
- 238000000855 fermentation Methods 0.000 claims description 36
- 230000004151 fermentation Effects 0.000 claims description 36
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 30
- 244000063299 Bacillus subtilis Species 0.000 claims description 27
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 27
- 239000000243 solution Substances 0.000 claims description 24
- 241000223602 Alternaria alternata Species 0.000 claims description 22
- 239000007788 liquid Substances 0.000 claims description 20
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 19
- 239000001963 growth medium Substances 0.000 claims description 18
- 238000009629 microbiological culture Methods 0.000 claims description 15
- 239000011780 sodium chloride Substances 0.000 claims description 15
- 238000009630 liquid culture Methods 0.000 claims description 14
- 230000004913 activation Effects 0.000 claims description 13
- 229920001817 Agar Polymers 0.000 claims description 12
- 239000008272 agar Substances 0.000 claims description 12
- 239000007787 solid Substances 0.000 claims description 12
- 238000012258 culturing Methods 0.000 claims description 11
- 238000000746 purification Methods 0.000 claims description 11
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 claims description 10
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 10
- 239000003761 preservation solution Substances 0.000 claims description 10
- 239000002609 medium Substances 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 7
- 238000003756 stirring Methods 0.000 claims description 6
- 235000019733 Fish meal Nutrition 0.000 claims description 5
- 235000019764 Soybean Meal Nutrition 0.000 claims description 5
- 229930006000 Sucrose Natural products 0.000 claims description 5
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 5
- 240000008042 Zea mays Species 0.000 claims description 5
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims description 5
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims description 5
- 239000002518 antifoaming agent Substances 0.000 claims description 5
- 229910000019 calcium carbonate Inorganic materials 0.000 claims description 5
- 235000005822 corn Nutrition 0.000 claims description 5
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 5
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019797 dipotassium phosphate Nutrition 0.000 claims description 5
- 239000004467 fishmeal Substances 0.000 claims description 5
- 235000013312 flour Nutrition 0.000 claims description 5
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 5
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 5
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 5
- 239000004455 soybean meal Substances 0.000 claims description 5
- 229960004793 sucrose Drugs 0.000 claims description 5
- 239000002054 inoculum Substances 0.000 claims description 3
- 238000000034 method Methods 0.000 claims description 3
- 238000002156 mixing Methods 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 24
- 230000008901 benefit Effects 0.000 abstract description 5
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 30
- 235000002637 Nicotiana tabacum Nutrition 0.000 description 28
- 201000010099 disease Diseases 0.000 description 27
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 27
- 241000208125 Nicotiana Species 0.000 description 24
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 20
- 230000001580 bacterial effect Effects 0.000 description 17
- 238000012360 testing method Methods 0.000 description 17
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 9
- 230000002265 prevention Effects 0.000 description 9
- 238000005507 spraying Methods 0.000 description 9
- 206010039509 Scab Diseases 0.000 description 8
- 238000013329 compounding Methods 0.000 description 6
- 230000003902 lesion Effects 0.000 description 6
- 244000052616 bacterial pathogen Species 0.000 description 5
- 239000011521 glass Substances 0.000 description 5
- 230000005764 inhibitory process Effects 0.000 description 5
- 238000010998 test method Methods 0.000 description 5
- 241000196324 Embryophyta Species 0.000 description 4
- 208000031888 Mycoses Diseases 0.000 description 4
- 244000061176 Nicotiana tabacum Species 0.000 description 4
- 230000000844 anti-bacterial effect Effects 0.000 description 4
- 239000003899 bactericide agent Substances 0.000 description 4
- 239000003085 diluting agent Substances 0.000 description 4
- 238000004321 preservation Methods 0.000 description 4
- 230000001954 sterilising effect Effects 0.000 description 4
- 238000004659 sterilization and disinfection Methods 0.000 description 4
- 241000233866 Fungi Species 0.000 description 3
- 230000003385 bacteriostatic effect Effects 0.000 description 3
- 238000001035 drying Methods 0.000 description 3
- 238000011835 investigation Methods 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 239000000575 pesticide Substances 0.000 description 3
- 238000007790 scraping Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 241001391944 Commicarpus scandens Species 0.000 description 2
- 239000000654 additive Substances 0.000 description 2
- 230000006806 disease prevention Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 239000003337 fertilizer Substances 0.000 description 2
- 244000053095 fungal pathogen Species 0.000 description 2
- 230000035784 germination Effects 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000000813 microbial effect Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000015097 nutrients Nutrition 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- 241000223600 Alternaria Species 0.000 description 1
- 208000035143 Bacterial infection Diseases 0.000 description 1
- 241000896246 Golovinomyces cichoracearum Species 0.000 description 1
- 229910002651 NO3 Inorganic materials 0.000 description 1
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 1
- 208000022362 bacterial infectious disease Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003124 biologic agent Substances 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 230000034303 cell budding Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 230000000003 effect on germination Effects 0.000 description 1
- 230000000459 effect on growth Effects 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 230000001678 irradiating effect Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000010979 pH adjustment Methods 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 1
Classifications
-
- 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
-
- 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
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
- A01N63/22—Bacillus
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Chemical & Material Sciences (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Organic Chemistry (AREA)
- Genetics & Genomics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Environmental Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Medicinal Chemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Pest Control & Pesticides (AREA)
- Biomedical Technology (AREA)
- Dentistry (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- Plant Pathology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The embodiment of the invention provides a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot, a preparation method and application thereof, and relates to the technical field of microorganisms. Has better control effect on powdery mildew and brown spot, can become endophytes of plants, has control effect for a long time, saves cost and improves economic benefit. And the preparation method of the bacillus compound microbial inoculum is simple and convenient, can be used for large-scale production, and is suitable for large-area popularization and application.
Description
Technical Field
The invention relates to the technical field of microorganisms, and particularly relates to a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot, and a preparation method and application thereof.
Background
The tobacco powdery mildew is a gas-fax bacterial disease caused by the infection of obligate parasitic fungus Erysiphe cichoracearum, and is commonly called as 'nitrate on the surface', 'grey on the surface', 'whitish' or 'frosting'. Since the discovery of tobacco powdery mildew in Taiwan province in 1919, tobacco powdery mildew was also discovered in Guizhou, Yunnan, Shanxi, Sichuan and Guangxi provinces in China. Tobacco powdery mildew can occur in the whole growth period of tobacco, mainly harms tobacco leaves, and can also spread to stems, flowering branches and sepals when serious. The damaged tobacco leaves are dark brown after being baked, lack elasticity and fragrance, and are easy to break, the loss is 10-20% in the light and more than 80% in the heavy, and serious yield reduction and quality reduction are caused. After the tobacco infected by the disease is modulated, the color is brown, obvious punctate disease spots exist on the leaf surface, the identity is thin, the oil content is less, the aroma is seriously deficient, the miscellaneous gas is heavy, the breakage degree is large, the crushing rate is high, the industrial availability is poor, and the economic value is lost.
The alternaria alternate disease is a leaf spot disease caused by alternaria fungus infection, and particularly becomes an obstacle factor influencing the maturity of tobacco leaves in the middle and later periods of tobacco leaf harvesting. The disease has the characteristics of short incubation period and high epidemic speed, and the tobacco plants are susceptible to the disease from budding to mature period, and can be epidemic in a large area in a short time under the condition of proper environmental conditions. Diseases begin to occur from leaves at the lower part of a tobacco plant, disease spots develop gradually from bottom to top along with the maturity of the leaves, the disease spots are fragile and easy to break, and when the diseases are serious, a plurality of disease spots are mutually connected and combined into large spots and scorched. In severe cases, stems, pedicels and capsules can be damaged, the yield and quality of tobacco leaves are affected, and the production of high-quality tobacco leaves is not facilitated, but no effective prevention and control means except chemical pesticides exist at present.
The powdery mildew and brown spot of tobacco are two main leaf fungal diseases in the tobacco production process, and the control of the two diseases is mainly dependent on chemical pesticides at present. Because the chemical pesticide control has the defects of pesticide residue, easy generation of resistance of pathogenic bacteria and the like, the quality of tobacco leaves is easy to reduce, the ecological environment is damaged, and the human health is influenced. Therefore, from the viewpoints of safety of tobacco leaves and environmental friendliness, the biological agent with the functions of preventing and treating powdery mildew and brown spot is particularly important for the green sustainable development of the tobacco industry.
Disclosure of Invention
The invention aims to provide a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and alternaria alternate, which can be used as a fungus bactericide, can be added with nutrients and additives to prepare a foliar fertilizer product with a disease prevention function for use, has a better prevention and treatment effect on the powdery mildew and the alternaria alternate, can become endophytes of plants, has a prevention and control effect for a long time, saves cost and improves economic benefits.
The second purpose of the invention is to provide a preparation method of the bacillus compound microbial inoculum, which is simple and convenient to operate and can be used for quickly and efficiently preparing the bacillus compound microbial inoculum.
The third purpose of the invention is to provide an application of the bacillus compound microbial inoculum, which can be used for preventing and treating powdery mildew and brown spot of tobacco and has higher application value.
The embodiment of the invention is realized by the following steps:
a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot comprises Bacillus belgii, Bacillus subtilis, a first Bacillus amyloliquefaciens and a second Bacillus amyloliquefaciens;
wherein the registration number of the Bacillus belgii in the China general microbiological culture Collection center is CGMCC No. 19886;
the registration number of the bacillus subtilis in China general microbiological culture Collection center is CGMCC No. 19887;
the registration number of the first bacillus amyloliquefaciens in the China general microbiological culture Collection center is CGMCC No. 19888;
the registration number of the second bacillus amyloliquefaciens in the China general microbiological culture Collection center is CGMCC No. 19889.
A preparation method of the bacillus compound microbial inoculum comprises the following steps:
respectively performing strain activation, strain purification and expanded culture on the preservation solution of the bacillus beleisis, the bacillus subtilis, the first bacillus amyloliquefaciens and the second bacillus amyloliquefaciens to obtain fermentation liquor of the bacillus beleisis, the bacillus subtilis, the first bacillus amyloliquefaciens and the second bacillus amyloliquefaciens;
and mixing fermentation liquids of the Bacillus belgii, the Bacillus subtilis, the first Bacillus amyloliquefaciens and the second Bacillus amyloliquefaciens to obtain the bacillus compound microbial inoculum.
An application of the bacillus compound microbial inoculum in treating powdery mildew and brown spot of tobacco.
The embodiment of the invention has the beneficial effects that:
the embodiment of the invention provides a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot, a preparation method and application thereof. Has better control effect on powdery mildew and brown spot, can become endophytes of plants, has control effect for a long time, saves cost and improves economic benefit. And the preparation method of the bacillus compound microbial inoculum is simple and convenient, can be used for large-scale production, and is suitable for large-area popularization and application.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
The bacillus compound microbial inoculum with the functions of preventing and treating powdery mildew and brown spot, the preparation method and the application thereof are concretely explained below.
The embodiment of the invention provides a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot, which comprises bacillus belgii, bacillus subtilis, a first bacillus amyloliquefaciens and a second bacillus amyloliquefaciens;
wherein the registration number of the Bacillus belgii in the China general microbiological culture Collection center is CGMCC No. 19886;
the registration number of the bacillus subtilis in China general microbiological culture Collection center is CGMCC No. 19887;
the registration number of the first bacillus amyloliquefaciens in the China general microbiological culture Collection center is CGMCC No. 19888;
the registration number of the second bacillus amyloliquefaciens in the China general microbiological culture Collection center is CGMCC No. 19889.
The four strains are carefully screened and compounded according to a specific proportion, so that the composition has an obvious prevention and treatment effect on powdery mildew and brown spot of tobacco.
Wherein, the Bacillus belgii is preserved in China general microbiological culture Collection center (CGMCC) in 5-month and 28-month 2020, and the address is as follows: china Beijing, the classification names are: bacillus belgii Bacillus velezensis with the preservation number: CGMCC No. 19886.
The bacillus subtilis is preserved in China general microbiological culture Collection center (CGMCC) in 5-month and 28-day 2020, and the address is as follows: china Beijing, the classification names are: the Bacillus subtilis has the following preservation number: CGMCC No. 19887.
The first bacillus amyloliquefaciens is preserved in China general microbiological culture Collection center (CGMCC) at 5-month and 28-month in 2020, and the address is as follows: china Beijing, the classification names are: bacillus amyloliquefaciens with a preservation number of: CGMCC No. 19888.
The second bacillus amyloliquefaciens is preserved in China general microbiological culture Collection center (CGMCC) at 5-month and 28-month in 2020, and the address is as follows: china Beijing, the classification names are: bacillus amyloliquefaciens with a preservation number of: CGMCC No. 19889.
In the bacillus compound microbial inoculum, the viable count ratio of the bacillus beleisi, the bacillus subtilis, the first bacillus amyloliquefaciens and the second bacillus amyloliquefaciens is 1: 2.8-3.2: 0.9-1.1: 2.8 to 3.2; preferably, the ratio of viable bacteria of bacillus belgii, bacillus subtilis, first bacillus amyloliquefaciens and second bacillus amyloliquefaciens is 1: 3: 1: 3; more preferably, the bacillusThe effective viable count of the bacterium compound microbial inoculum is more than 2.0 × 1010cfu/g. The bacillus complex inoculant has better control effect according to the proportion.
The embodiment of the invention also provides a preparation method of the bacillus compound microbial inoculum, which comprises the following steps:
s1, respectively carrying out strain activation, strain purification and expanded culture on the preservation solution of the Bacillus belgii, the Bacillus subtilis, the first Bacillus amyloliquefaciens and the second Bacillus amyloliquefaciens to obtain fermentation liquor of the Bacillus belgii, the Bacillus subtilis, the first Bacillus amyloliquefaciens and the second Bacillus amyloliquefaciens;
and S2, mixing fermentation liquids of the Bacillus belgii, the Bacillus subtilis, the first Bacillus amyloliquefaciens and the second Bacillus amyloliquefaciens to obtain the bacillus compound microbial inoculum.
The strain activation is to inoculate the preservation solution to a beef extract peptone liquid culture medium, and culture the beef extract peptone liquid culture medium for 48-60 hours at the temperature of 33-37 ℃ and the stirring speed of 150-180 rpm to obtain an activation culture solution; the volume ratio of the preservation solution to the beef extract peptone liquid medium is 1: 90-120 parts. Wherein, according to the parts by weight, the beef extract peptone liquid medium comprises: 2-4 parts of beef extract, 8-11 parts of peptone, 4-6 parts of sodium chloride and 900-1000 parts of water; the pH value of the beef extract peptone liquid culture medium is 6-8.
And the strain purification is to inoculate the activated culture solution to a beef extract peptone solid plate for streak culture, culture for 72-80 h at 33-37 ℃, and select a single larger bacterial colony with the bacterial colony characteristics in accordance with as the activated seed bacterial colony. Wherein, according to parts by weight, the beef extract peptone solid flat board includes: 2-4 parts of beef extract, 8-11 parts of peptone, 4-6 parts of sodium chloride, 15-25 parts of agar and 900-1000 parts of water; the pH value of the beef extract peptone solid flat plate is 6-8.
The expanding culture is to inoculate the seed bacterial colony into a beef peptone liquid culture medium, and culture the seed bacterial colony for 60-72 hours at the temperature of 33-37 ℃ and the stirring speed of 150-180 rpm to obtain a seed solution; and inoculating the seed liquid into the fermentation culture liquid according to the proportion of 2-2.5 per mill, and culturing for 36-48 h at the temperature of 33-37 ℃ and the stirring speed of 100-150 rpm to obtain the fermentation liquid. Wherein, according to the weight portion, the fermentation culture solution comprises: 2-4 parts of soybean meal, 0.5-1 part of cane sugar, 1-3 parts of beef extract, 2-3 parts of corn flour, 0.8-1.5 parts of calcium carbonate, 1-2 parts of fish meal, 0.05-0.1 part of monopotassium phosphate, 0.05-0.1 part of dipotassium phosphate, 1-5 parts of sodium chloride and 0.1-0.3 part of defoaming agent; the pH value of the fermentation culture solution is 6.8-7.5.
The preparation method is simple to operate, and can be used for quickly and efficiently culturing the required fermentation liquor. It should be noted that, in order to avoid introducing bacteria, all the transfer operations in the above steps are performed using a sterile inoculating loop. Further, pH adjustment for each medium was performed using hydrochloric acid and sodium hydroxide.
The embodiment of the invention also provides an application of the bacillus compound microbial inoculum in treating powdery mildew and brown spot of tobacco.
The features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The embodiment provides a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot, which comprises the following preparation steps:
s1, strain activation: respectively inoculating 0.1 ml of preservation solution of bacillus belgii, bacillus subtilis, first bacillus amyloliquefaciens and second bacillus amyloliquefaciens which are preserved by adopting glycerol at the temperature of-80 ℃ into 10 ml of beef extract peptone liquid culture medium, and carrying out activated culture for 48 h under the conditions that the temperature is 35 ℃ and the rotating speed is 160 rpm to obtain activated culture solution of four strains;
the beef extract peptone liquid culture medium comprises the following components in parts by weight: 3.0 g of beef extract, 10.0 g of peptone, 5.0 g of sodium chloride and 1000 ml of water, and adjusting the pH value to 6-8 by adopting hydrochloric acid and sodium hydroxide.
S2, strain purification: respectively inoculating strains in the activation culture solution to a beef extract peptone solid plate by using a sterilization inoculating loop, carrying out purification streak culture, culturing for 3 days in an incubator at the temperature of 35 ℃, and selecting a single larger bacterial colony with the bacterial colony characteristics conforming to the bacterial colony characteristics of the functional strains as an activated seed bacterial colony;
the beef extract peptone solid flat plate comprises the following components in parts by weight: 3.0 g of beef extract, 10.0 g of peptone, 5.0 g of sodium chloride, 25 g of agar and 1000 ml of water, and the pH is adjusted to be between 6 and 8 by adopting hydrochloric acid and sodium hydroxide.
S3, enlarged culture: scraping the activated seed colonies in the first step in one loop by using an aseptic inoculating loop, inoculating the seed colonies into 50 ml of beef extract peptone liquid culture medium, and performing activated culture for 72 h under the conditions that the temperature is 35 ℃ and the rotating speed is 160 rpm to obtain seed liquid of four strains; inoculating the strain seed liquid into a fermentation culture solution in a fermentation tank according to the proportion of 2 per mill, and culturing for 36 h under the conditions that the temperature is 35 ℃ and the rotating speed is 120 rpm to obtain fermentation liquor of four strains;
wherein the fermentation culture solution comprises the following components in parts by weight: 2 parts of soybean meal, 1 part of cane sugar, 1 part of beef extract, 3 parts of corn flour, 0.8 part of calcium carbonate, 2 parts of fish meal, 0.05 part of monopotassium phosphate, 0.1 part of dipotassium phosphate, 1 part of sodium chloride and 0.3 part of defoaming agent. Adjusting the pH value to 6.8-7.5 by using hydrochloric acid and sodium hydroxide.
S4: compounding: fermenting the fermentation liquor of Bacillus belgii, Bacillus subtilis, first Bacillus amyloliquefaciens and second Bacillus amyloliquefaciens according to the viable count ratio of 1: 3: 1: 3, compounding to obtain the required bacillus compound microbial inoculum.
Example 2
The embodiment provides a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot, which comprises the following preparation steps:
s1, strain activation: respectively inoculating 0.1 ml of preservation solution of bacillus belgii, bacillus subtilis, first bacillus amyloliquefaciens and second bacillus amyloliquefaciens which are preserved by adopting glycerol at the temperature of-80 ℃ into 10 ml of beef extract peptone liquid culture medium, and carrying out activated culture for 60 h under the conditions that the temperature is 37 ℃ and the rotating speed is 180rpm to obtain activated culture solution of four strains;
the beef extract peptone liquid culture medium comprises the following components in parts by weight: 4.0 g of beef extract, 8.0 g of peptone, 6.0 g of sodium chloride and 900 ml of water, and adjusting the pH value to 6-8 by adopting hydrochloric acid and sodium hydroxide.
S2, strain purification: respectively inoculating strains in the activation culture solution to a beef extract peptone solid plate by using a sterilization inoculating loop, carrying out purification streak culture, culturing for 80 h in an incubator at the temperature of 37 ℃, and selecting a single larger bacterial colony with the bacterial colony characteristics conforming to the bacterial colony characteristics of the functional strains as an activated seed bacterial colony;
the beef extract peptone solid flat plate comprises the following components in parts by weight: 4.0 g of beef extract, 8.0 g of peptone, 6.0 g of sodium chloride, 20 g of agar and 900 ml of water, and the pH is adjusted to be between 6 and 8 by adopting hydrochloric acid and sodium hydroxide.
S3, enlarged culture: scraping the activated seed colonies in the first step in one loop by using an aseptic inoculating loop, inoculating the seed colonies into 50 ml of beef extract peptone liquid culture medium, and performing activated culture for 60 hours under the conditions that the temperature is 37 ℃ and the rotating speed is 180rpm to obtain seed liquid of four strains; inoculating the strain seed liquid into a fermentation culture solution in a fermentation tank according to the proportion of 2.5 per mill, and culturing for 48 h under the conditions that the temperature is 37 ℃ and the rotating speed is 150 rpm to obtain fermentation liquor of four strains;
wherein the fermentation culture solution comprises the following components in parts by weight: 4 parts of soybean meal, 0.5 part of cane sugar, 3 parts of beef extract, 2 parts of corn flour, 1.5 parts of calcium carbonate, 1 part of fish meal, 0.1 part of monopotassium phosphate, 0.05 part of dipotassium phosphate, 3 parts of sodium chloride and 0.1 part of defoaming agent. Adjusting the pH value to 6.8-7.5 by using hydrochloric acid and sodium hydroxide.
S4: compounding: fermenting the fermentation liquor of Bacillus belgii, Bacillus subtilis, first Bacillus amyloliquefaciens and second Bacillus amyloliquefaciens according to the viable count ratio of 1: 2.8: 1.1: 3.2, compounding to obtain the required bacillus compound microbial inoculum.
Example 3
The embodiment provides a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot, which comprises the following preparation steps:
s1, strain activation: respectively inoculating 0.1 ml of preservation solution of bacillus belgii, bacillus subtilis, first bacillus amyloliquefaciens and second bacillus amyloliquefaciens which are preserved by adopting glycerol at the temperature of-80 ℃ into 10 ml of beef extract peptone liquid culture medium, and carrying out activated culture for 48 h under the conditions that the temperature is 33 ℃ and the rotating speed is 150 rpm to obtain activated culture solution of four strains;
the beef extract peptone liquid culture medium comprises the following components in parts by weight: 2.0 g of beef extract, 11.0 g of peptone, 4.0 g of sodium chloride and 1000 ml of water, and adjusting the pH value to 6-8 by adopting hydrochloric acid and sodium hydroxide.
S2, strain purification: respectively inoculating strains in the activation culture solution to a beef extract peptone solid plate by using a sterilization inoculating loop, carrying out purification streak culture, culturing for 72 h in an incubator at the temperature of 34 ℃, and selecting a single larger bacterial colony with the bacterial colony characteristics conforming to the bacterial colony characteristics of the functional strains as an activated seed bacterial colony;
the beef extract peptone solid flat plate comprises the following components in parts by weight: 2.0 g of beef extract, 11.0 g of peptone, 4.0 g of sodium chloride, 23 g of agar and 1000 ml of water, and the pH is adjusted to be between 6 and 8 by adopting hydrochloric acid and sodium hydroxide.
S3, enlarged culture: scraping the activated seed colonies in the first step in one loop by using an aseptic inoculating loop, inoculating the seed colonies into 50 ml of beef extract peptone liquid culture medium, and performing activated culture for 72 h under the conditions that the temperature is 34 ℃ and the rotating speed is 150 rpm to obtain seed liquid of four strains; inoculating the strain seed liquid into a fermentation culture solution in a fermentation tank according to the proportion of 2 per mill, and culturing for 36 h under the conditions that the temperature is 34 ℃ and the rotating speed is 100 rpm to obtain fermentation liquor of four strains;
wherein the fermentation culture solution comprises the following components in parts by weight: 2 parts of soybean meal, 1 part of cane sugar, 2 parts of beef extract, 2 parts of corn flour, 1 part of calcium carbonate, 1.5 parts of fish meal, 0.08 part of monopotassium phosphate, 0.06 part of dipotassium phosphate, 5 parts of sodium chloride and 0.2 part of defoaming agent. Adjusting the pH value to 6.8-7.5 by using hydrochloric acid and sodium hydroxide.
S4: compounding: fermenting the fermentation liquor of Bacillus belgii, Bacillus subtilis, first Bacillus amyloliquefaciens and second Bacillus amyloliquefaciens according to the viable count ratio of 1: 3.2: 0.9: 2.8, compounding to obtain the required bacillus compound microbial inoculum.
Test example 1
The bacillus compound microbial inoculum provided by the embodiment 1-3 is used for testing the inhibition effect on the powdery mildew and the brown spot of tobacco, and the specific test method is as follows:
1. test site:
laboratory of research center for microbial fermentation engineering, Yunnan, Anning, Yunnan, China.
2. Test materials
Test medium: PSA culture medium (activated tobacco brown spot pathogenic bacteria), LB culture medium (culture function compound bacteria agent), and 2% water agar culture medium.
Test materials Bacillus Complex microbial inoculum (diluted with water to effective viable count of about 1 × 10) of examples 1-37cfu/mL), powdery mildew, scab pathogens.
3. Test method
Powdery mildew of tobacco: preparing a 2% water agar culture medium, pouring the water agar on a clean glass slide in a super clean workbench after sterilization to form a thin layer, irradiating an ultraviolet lamp, and cooling for later use. 0.15 ml of clear water and the bacillus compound bactericide are respectively absorbed and added onto an agar glass, powdery mildew is scraped and spread on the agar glass to be mixed with the compound bactericide or clear water, the mixture is coated on the agar glass, then the agar glass is placed in a sterile culture dish with a dish bottom and a dish cover both stuck with moist filter paper, the culture dish is placed in a 25 ℃ fungus incubator to be subjected to dark culture for 24 hours and then subjected to microscopic examination, 1/2 with the length of a germ tube exceeding the short diameter of the spore is regarded as germination, 150 spores are microscopically examined at each concentration, and each slide is microscopically examined twice.
Alternaria alternate: 4 points 3 cm away from the center on the PSA culture plate coated with the alternaria alternate pathogenic fungi are inoculated with a functional compound microbial inoculum, only the alternaria alternate pathogenic fungi are coated for comparison, the culture plate is cultured in a dark mode until the control plate alternaria alternate fungi grow over the whole dish, the bacteriostatic zone is measured and recorded, and 5 plates are repeated for each treatment. The bacteriostatic rate (%) (control colony diameter-treated colony diameter ÷ control colony diameter ] × 100.
The test results are shown in tables 1 and 2.
TABLE 1 measurement of control Effect of powdery mildewTest for
| Treatment of | Average number of germinating spores | Microscopic examination of total number of spores | Germination Rate (%) | Inhibition ratio (%) |
| Clean water | 32.5 | 150 | 21.67 | - |
| Example 1 | 13.5 | 150 | 9.00 | 58.46 |
| Example 2 | 15.5 | 150 | 10.33 | 52.33 |
| Example 3 | 16.0 | 150 | 10.67 | 50.76 |
TABLE 2 test of prevention and treatment effects on Alternaria alternate
| Treatment of | Average width of bacteriostatic zone/mm | Inhibition rate/%) |
| Control group | - | - |
| Example 1 | 10.5±0.58 | 56.70±1.27 |
| Example 2 | 8.3±0.42 | 44.86±1.35 |
| Example 3 | 7.9±0.46 | 42.70±1.25 |
As can be seen from tables 1 and 2, the bacillus complex microbial inoculum provided by embodiments 1 to 3 of the present invention has an obvious inhibition effect on germination of powdery mildew and growth of brown spot hyphae, and it can be intuitively known that the bacillus complex microbial inoculum has a good control effect on tobacco powdery mildew and brown spot, has a good inhibition effect on growth and propagation of pathogenic fungi, and has a good control effect on fungal diseases.
Test example 2
The bacillus compound microbial inoculum provided by the embodiment 1 is used for testing the prevention and control effect of the bacillus compound microbial inoculum on the powdery mildew and the brown spot of tobacco in the growth process of the tobacco, and the specific test method is as follows:
1 basic conditions of the test
The test is carried out in a test greenhouse of the research center of microbial fermentation engineering, GmbH, Yunnan, Qinglongzhen, Anning, Yunnan, province.
2 test materials
4-5 true-leaf Yunyan 87, the bacillus compound microbial inoculum provided by the example 1, pure fermentation liquor (no inoculation), and spore liquid of powdery mildew and brown spot pathogenic bacteria are cultured in a small pot plant. Diluting the functional compound microbial inoculum and the pure fermentation liquor by 200 times for later use.
3 test method
3.1 design of the experiment
In the experiment, the cloud 87 tobacco leaves are treated, 4 times of treatment are set, and the treatment is as follows:
and (3) controlling powdery mildew:
treatment 1: spraying 10 mL of clear water, drying, and inoculating powdery mildew pathogenic bacteria spores
And (3) treatment 2: spraying 10 mL of pure fermentation broth diluent, drying, and inoculating powdery mildew pathogenic bacteria spores
And (3) treatment: spraying 10 mL of bacillus compound microbial inoculum diluent, drying, and inoculating powdery mildew pathogenic spore
Prevention and treatment of brown spot: and (3) inoculating the alternaria alternate spores to the tobacco leaves, recording the number of the alternaria alternate lesions on each leaf when the tobacco leaves have the lesions, and then processing.
And (4) treatment: spraying 10 mL of clear water
And (4) treatment 5: spraying 10 mL of pure fermentation broth diluent
And (6) treatment: spraying 10 mL of bacillus compound microbial inoculum diluent
The effective viable count of the compound microbial inoculum is 1.0 × 107cfu/mL。
3.2 test procedure
And screening out tobacco plants with similar growth vigor for testing. According to the treatment arrangement test, in the control of powdery mildew, 7 d, 14 d and 21 d after inoculation are respectively used for investigating the powdery mildew disease incidence condition of each treatment according to the national tobacco powdery mildew disease severity grading standard, and calculating the disease index and the relative control effect; in the control of the alternaria alternate, after treatment, spraying is carried out again at intervals of 5 days, the spraying is carried out for 3 times totally, and the control effect investigation is carried out on the 5 th day after the spraying respectively.
3.3 investigation method
Powdery mildew is graded and investigated by taking leaves as a unit:
level 0: no scab
Level 1: the area of the lesion spots accounts for less than 5% of the area of the leaf.
And 3, level: the area of the scab accounts for 6-10% of the area of the leaf.
And 5, stage: the area of the scab accounts for 11-20% of the area of the leaf.
And 7, stage: the area of the scab accounts for 21-40% of the area of the leaf.
And 9, stage: the lesion area accounts for more than 41% of the leaf area.
Disease index = (number of diseased leaves at each stage × disease representative value) × 100/(survey total number of leaves × 9)
Control effect (%) [ (control disease index-treatment disease index)/control disease index ] × 100
Red star disease: the leaf blade is used as a unit for grading investigation.
Level 0: disease of whole leaf
Level 1: the area of the lesion spots accounts for less than 1% of the area of the leaves.
And 3, level: the area of the scab accounts for 2 to 5 percent of the area of the leaf.
And 5, stage: the area of the scab accounts for 6-10% of the area of the leaf.
And 7, stage: the area of the scab accounts for 11-20% of the area of the leaf.
And 9, stage: the area of the lesion spots accounts for more than 21 percent of the area of the leaves.
Disease index = (number of diseased leaves at each stage × disease representative value) × 100/(survey total number of leaves × 9)
Control effect (%) [ (control disease index-treatment disease index)/control disease index ] × 100
The test results are shown in tables 3 and 4.
TABLE 3 control of powdery mildew
TABLE 4 prevention and treatment of Alternaria alternate
As can be seen from tables 3 and 4, the relative control effect of the bacillus compound microbial inoculum provided by the embodiment of the invention on powdery mildew can reach 83.28% within 7 days, and 45.54% can still be controlled after 21 days. Meanwhile, the control effect on the alternaria alternate can reach 74.2% -79.4% in 5 days after each administration. Fully shows that the bacillus compound microbial inoculum of the embodiment of the invention can play a certain prevention and control effect on powdery mildew and brown spot of tobacco, can effectively inhibit the growth of pathogenic fungi, prevents and controls fungal diseases, and has good effect on improving the quality of tobacco leaves.
In conclusion, the embodiment of the invention provides a bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot, a preparation method and application thereof, the compound microbial inoculum is prepared by bacillus strains carefully selected by an inventor according to a specific proportion, and the compound microbial inoculum can be directly used as a fungus bactericide and can also be added with nutrients and additives to prepare a foliar fertilizer product with a disease prevention function for use. Has better control effect on powdery mildew and brown spot, can become endophytes of plants, has control effect for a long time, saves cost and improves economic benefit. And the preparation method of the bacillus compound microbial inoculum is simple and convenient, can be used for large-scale production, and is suitable for large-area popularization and application.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
1. A bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot is characterized by comprising Bacillus belgii, Bacillus subtilis, a first Bacillus amyloliquefaciens and a second Bacillus amyloliquefaciens;
wherein the registration number of the Bacillus belgii in the China general microbiological culture Collection center is CGMCC No. 19886;
the registration number of the bacillus subtilis in the China general microbiological culture Collection center is CGMCC No. 19887;
the registration number of the first bacillus amyloliquefaciens in the China general microbiological culture Collection center is CGMCC No. 19888;
the registration number of the second bacillus amyloliquefaciens in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No. 19889.
2. The bacillus complex inoculant according to claim 1, wherein the ratio of viable bacteria of the bacillus belief, the bacillus subtilis, the first bacillus amyloliquefaciens and the second bacillus amyloliquefaciens is equal to1: 2.8-3.2: 0.9-1.1: 2.8-3.2, preferably the viable bacteria ratio of the Bacillus belgii, the Bacillus subtilis, the first Bacillus amyloliquefaciens and the second Bacillus amyloliquefaciens is 1: 3: 1: 3, and preferably the effective viable bacteria number of the bacillus compound microbial inoculum is more than 2.0 × 1010cfu/g。
3. The preparation method of the bacillus compound microbial inoculum of claim 1 or 2, which comprises the following steps:
respectively performing strain activation, strain purification and expanded culture on the preservation solution of the bacillus belgii, the bacillus subtilis, the first bacillus amyloliquefaciens and the second bacillus amyloliquefaciens to obtain fermentation liquor of the bacillus belgii, the bacillus subtilis, the first bacillus amyloliquefaciens and the second bacillus amyloliquefaciens;
and mixing the fermentation liquor of the Bacillus belgii, the Bacillus subtilis, the first Bacillus amyloliquefaciens and the second Bacillus amyloliquefaciens to obtain the bacillus compound microbial inoculum.
4. The preparation method according to claim 3, wherein the strain activation is carried out by inoculating the preservation solution to a beef extract peptone liquid medium, and culturing at 33-37 ℃ and a stirring speed of 150-180 rpm for 48-60 h to obtain an activation culture solution; the volume ratio of the preservation solution to the beef extract peptone liquid medium is 1: 90-120 parts.
5. The preparation method according to claim 4, wherein the beef extract peptone liquid medium comprises, in parts by weight: 2-4 parts of beef extract, 8-11 parts of peptone, 4-6 parts of sodium chloride and 900-1000 parts of water; the pH value of the beef extract peptone liquid culture medium is 6-8.
6. The preparation method according to claim 4, wherein the strain purification comprises inoculating the activated culture solution to a beef extract peptone solid plate for streak culture, culturing at 33-37 ℃ for 72-80 h, and selecting a single larger colony with a consistent colony characteristic as an activated seed colony.
7. The preparation method according to claim 6, wherein the beef extract peptone solid tablet comprises, in parts by weight: 2-4 parts of beef extract, 8-11 parts of peptone, 4-6 parts of sodium chloride, 15-25 parts of agar and 900-1000 parts of water; the pH value of the beef extract peptone solid flat plate is 6-8.
8. The preparation method according to claim 6, wherein the expanded culture is to inoculate the seed colonies into the beef peptone liquid medium, and culture the seed colonies for 60-72 hours at 33-37 ℃ and at a stirring speed of 150-180 rpm to obtain a seed solution; and inoculating the seed liquid into a fermentation culture solution according to the proportion of 2-2.5 per mill, and culturing for 36-48 h at the temperature of 33-37 ℃ and the stirring speed of 100-150 rpm to obtain the fermentation liquid.
9. The method according to claim 8, wherein the fermentation broth comprises, in parts by weight: 2-4 parts of soybean meal, 0.5-1 part of cane sugar, 1-3 parts of beef extract, 2-3 parts of corn flour, 0.8-1.5 parts of calcium carbonate, 1-2 parts of fish meal, 0.05-0.1 part of monopotassium phosphate, 0.05-0.1 part of dipotassium phosphate, 1-5 parts of sodium chloride and 0.1-0.3 part of defoaming agent; the pH value of the fermentation culture solution is 6.8-7.5.
10. The application of the bacillus complex inoculant as defined in claim 1 or 2 in treating powdery mildew and brown spot of tobacco.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010652997.4A CN111748498A (en) | 2020-07-08 | 2020-07-08 | Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010652997.4A CN111748498A (en) | 2020-07-08 | 2020-07-08 | Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111748498A true CN111748498A (en) | 2020-10-09 |
Family
ID=72711059
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010652997.4A Pending CN111748498A (en) | 2020-07-08 | 2020-07-08 | Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111748498A (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468057A (en) * | 2019-09-02 | 2019-11-19 | 昆明理工大学 | One plants endogenetic disk both-end hair spore category fungi M7SB 41 and its application |
| CN112322536A (en) * | 2020-11-11 | 2021-02-05 | 云南省烟草农业科学研究院 | Bacillus belgii Z002, and acquisition method and application thereof |
| CN116590170A (en) * | 2022-12-26 | 2023-08-15 | 安徽科技学院 | Preparation and application of microorganism strain, compound strain and growth-promoting disease-preventing organic fertilizer |
Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101423812A (en) * | 2008-12-17 | 2009-05-06 | 河南省农业科学院 | Bacillus amyloliquefaciens and microbiological preparation and preparation method thereof |
| CN103695356A (en) * | 2013-12-24 | 2014-04-02 | 河北省科学院生物研究所 | Bacillus amyloliquefaciens, microbial inoculant and application thereof |
| CN105733991A (en) * | 2016-03-30 | 2016-07-06 | 贵州大学 | Bacillus amyloliquefaciens strain and application thereof in preventing and controlling powdery mildew of tobacco |
| CN108641981A (en) * | 2018-05-17 | 2018-10-12 | 十堰市农业科学院 | One plant of Biocontrol Bacillus identification and application |
| CN108753664A (en) * | 2018-06-28 | 2018-11-06 | 广东省农业科学院农业资源与环境研究所 | A kind of biocontrol agent, biological organic fertilizer and its application |
| CN109699682A (en) * | 2019-03-07 | 2019-05-03 | 东北农业大学 | A kind of tomato biological and ecological methods to prevent plant disease, pests, and erosion seed coat agent and its application |
| CN110295129A (en) * | 2019-07-23 | 2019-10-01 | 山东省农药科学研究院 | A kind of biocontrol microorganisms and application for preventing and treating gray mold of cucumber and powdery mildew |
| CN110402964A (en) * | 2019-06-28 | 2019-11-05 | 中国烟草中南农业试验站 | A kind of microbial bacterial agent and preparation method thereof for preventing and treating Alternaria alternate |
| CN110564655A (en) * | 2019-10-17 | 2019-12-13 | 中国烟草总公司陕西省公司 | Preparation and application of bacillus subtilis SL-19 microbial inoculum for biological control of alternaria alternate |
| CN110938573A (en) * | 2019-12-20 | 2020-03-31 | 云南省微生物发酵工程研究中心有限公司 | Preparation method of bacillus subtilis 24-7 strain fermentation liquid |
| CN111100820A (en) * | 2020-01-15 | 2020-05-05 | 河北省农林科学院植物保护研究所 | Bacillus belgii HMB28023 and application thereof |
| JP2020079269A (en) * | 2020-01-31 | 2020-05-28 | 住友化学株式会社 | Plant disease control method |
| CN111286473A (en) * | 2019-12-20 | 2020-06-16 | 云南省微生物发酵工程研究中心有限公司 | Agricultural microbial bacterium composite inoculant and preparation method thereof |
| CN111411063A (en) * | 2020-05-15 | 2020-07-14 | 云南省微生物发酵工程研究中心有限公司 | Bacillus composite microbial inoculum with function of preventing and treating tobacco soil-borne diseases and preparation method thereof |
| CN111747789A (en) * | 2020-07-08 | 2020-10-09 | 云南省烟草公司曲靖市公司 | Biological foliar fertilizer with functions of preventing and treating powdery mildew and alternaria alternate, and preparation method and application thereof |
| CN112358997A (en) * | 2020-11-20 | 2021-02-12 | 云南省烟草公司曲靖市公司 | Bacterial strain with biocontrol function on tobacco brown spot, fermentation method of bacterial strain and tobacco brown spot prevention and control compound agent |
| CN112391315A (en) * | 2020-11-20 | 2021-02-23 | 中国农业科学院烟草研究所 | Bacillus belgii and microbial inoculum and application thereof |
| CN114478109A (en) * | 2021-12-27 | 2022-05-13 | 云南云叶化肥股份有限公司 | Biological foliar fertilizer and preparation method thereof |
-
2020
- 2020-07-08 CN CN202010652997.4A patent/CN111748498A/en active Pending
Patent Citations (18)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101423812A (en) * | 2008-12-17 | 2009-05-06 | 河南省农业科学院 | Bacillus amyloliquefaciens and microbiological preparation and preparation method thereof |
| CN103695356A (en) * | 2013-12-24 | 2014-04-02 | 河北省科学院生物研究所 | Bacillus amyloliquefaciens, microbial inoculant and application thereof |
| CN105733991A (en) * | 2016-03-30 | 2016-07-06 | 贵州大学 | Bacillus amyloliquefaciens strain and application thereof in preventing and controlling powdery mildew of tobacco |
| CN108641981A (en) * | 2018-05-17 | 2018-10-12 | 十堰市农业科学院 | One plant of Biocontrol Bacillus identification and application |
| CN108753664A (en) * | 2018-06-28 | 2018-11-06 | 广东省农业科学院农业资源与环境研究所 | A kind of biocontrol agent, biological organic fertilizer and its application |
| CN109699682A (en) * | 2019-03-07 | 2019-05-03 | 东北农业大学 | A kind of tomato biological and ecological methods to prevent plant disease, pests, and erosion seed coat agent and its application |
| CN110402964A (en) * | 2019-06-28 | 2019-11-05 | 中国烟草中南农业试验站 | A kind of microbial bacterial agent and preparation method thereof for preventing and treating Alternaria alternate |
| CN110295129A (en) * | 2019-07-23 | 2019-10-01 | 山东省农药科学研究院 | A kind of biocontrol microorganisms and application for preventing and treating gray mold of cucumber and powdery mildew |
| CN110564655A (en) * | 2019-10-17 | 2019-12-13 | 中国烟草总公司陕西省公司 | Preparation and application of bacillus subtilis SL-19 microbial inoculum for biological control of alternaria alternate |
| CN110938573A (en) * | 2019-12-20 | 2020-03-31 | 云南省微生物发酵工程研究中心有限公司 | Preparation method of bacillus subtilis 24-7 strain fermentation liquid |
| CN111286473A (en) * | 2019-12-20 | 2020-06-16 | 云南省微生物发酵工程研究中心有限公司 | Agricultural microbial bacterium composite inoculant and preparation method thereof |
| CN111100820A (en) * | 2020-01-15 | 2020-05-05 | 河北省农林科学院植物保护研究所 | Bacillus belgii HMB28023 and application thereof |
| JP2020079269A (en) * | 2020-01-31 | 2020-05-28 | 住友化学株式会社 | Plant disease control method |
| CN111411063A (en) * | 2020-05-15 | 2020-07-14 | 云南省微生物发酵工程研究中心有限公司 | Bacillus composite microbial inoculum with function of preventing and treating tobacco soil-borne diseases and preparation method thereof |
| CN111747789A (en) * | 2020-07-08 | 2020-10-09 | 云南省烟草公司曲靖市公司 | Biological foliar fertilizer with functions of preventing and treating powdery mildew and alternaria alternate, and preparation method and application thereof |
| CN112358997A (en) * | 2020-11-20 | 2021-02-12 | 云南省烟草公司曲靖市公司 | Bacterial strain with biocontrol function on tobacco brown spot, fermentation method of bacterial strain and tobacco brown spot prevention and control compound agent |
| CN112391315A (en) * | 2020-11-20 | 2021-02-23 | 中国农业科学院烟草研究所 | Bacillus belgii and microbial inoculum and application thereof |
| CN114478109A (en) * | 2021-12-27 | 2022-05-13 | 云南云叶化肥股份有限公司 | Biological foliar fertilizer and preparation method thereof |
Non-Patent Citations (7)
| Title |
|---|
| BEN FAN等: "Bacillus amyloliquefaciens, Bacillus velezensis, and Bacillus siamensis Form an "Operational Group B. amyloliquefaciens" within the B. subtilis Species Complex", 《FRONT MICROBIOL》 * |
| DEBORAH R. FRAVEL等: "Biocontrol of Tobacco Brown-Spot Disease by Bacillus cereus subsp. mycoides in a Controlled Environment", 《DISEASE CONTROL AND PEST MANAGEMENT》 * |
| RONG JIAO等: "Bacillus amyloliquefaciens YN201732 Produces Lipopeptides With Promising Biocontrol Activity Against Fungal Pathogen Erysiphe cichoracearum", 《FRONT CELL INFECT MICROBIOL》 * |
| YOUNG-SOOK KIM等: "Bacillus sp. BS061 Suppresses Powdery Mildew and Gray Mold", 《 MYCOBIOLOGY》 * |
| 司世飞: "烟草赤星病生防菌筛选及发酵条件优化", 《中国优秀博硕士学位论文全文数据库(硕士)农业科技辑（电子期刊）》 * |
| 武君洁等: "枯草芽胞杆菌YN2014090悬浮剂的研制", 《中国生物防治学报》 * |
| 焦蓉等: "内生菌YN201728的定殖能力及其防治烟草白粉病的效果研究", 《核农学报》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110468057A (en) * | 2019-09-02 | 2019-11-19 | 昆明理工大学 | One plants endogenetic disk both-end hair spore category fungi M7SB 41 and its application |
| CN110468057B (en) * | 2019-09-02 | 2021-09-24 | 昆明理工大学 | Endophytic pestalotiopsis fungus M7SB41 and application thereof |
| CN112322536A (en) * | 2020-11-11 | 2021-02-05 | 云南省烟草农业科学研究院 | Bacillus belgii Z002, and acquisition method and application thereof |
| CN112322536B (en) * | 2020-11-11 | 2024-07-16 | 云南省烟草农业科学研究院 | Bacillus bailii Z002, acquisition method and application thereof |
| CN116590170A (en) * | 2022-12-26 | 2023-08-15 | 安徽科技学院 | Preparation and application of microorganism strain, compound strain and growth-promoting disease-preventing organic fertilizer |
| CN116590170B (en) * | 2022-12-26 | 2024-02-06 | 安徽科技学院 | Preparation and application of microorganism strain, compound strain and growth-promoting disease-preventing organic fertilizer |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111500501B (en) | Streptomyces misonii strain and application thereof in preventing and treating wheat root rot and stem basal rot | |
| CN111748498A (en) | Bacillus compound microbial inoculum with functions of preventing and treating powdery mildew and brown spot as well as preparation method and application thereof | |
| CN114854627B (en) | Pseudomonas fluorescens for preventing and treating bacterial wilt and application thereof | |
| CN113980846B (en) | High-efficiency stress-resistant bacillus subtilis for antagonizing fusarium oxysporum | |
| CN109456921B (en) | Paenibacillus polymyxa, application thereof, microbial agent, powder and granules | |
| CN114875016B (en) | Formulated carrier suitable for pseudomonas fluorescens and microbial inoculum thereof | |
| CN110317747B (en) | Bacillus amyloliquefaciens JT68 and application thereof in prevention and treatment of tea anthracnose | |
| CN113215016B (en) | Bacillus amyloliquefaciens and application thereof | |
| CN113549578B (en) | Bacillus siamensis BsNlG13 for inhibiting Pyricularia oryzae and promoting seed germination and application thereof | |
| CN112342173B (en) | Bacillus belgii and application thereof | |
| CN111172063A (en) | Bacillus beleisi MES861 and application and product thereof | |
| CN108641989A (en) | One plant of Methylotrophic bacillus and its application | |
| CN110396485B (en) | Brevibacillus brevis for producing auxin and application thereof | |
| CN116731892A (en) | Antibacterial bacillus bailii Y103-16 and application thereof | |
| CN109294961B (en) | Biocontrol strain PNC25 for preventing and treating litchi downy blight and application thereof | |
| CN117126790B (en) | Serratia pratensis and application thereof in inhibition of zearalenone | |
| CN111662843B (en) | Bacillus amyloliquefaciens and separation and screening method and application thereof | |
| CN115851479B (en) | Bacteria with antagonism to botrytis cinerea and application thereof | |
| CN116590199A (en) | Paenibacillus piri and application thereof in prevention and treatment of corn ear rot | |
| CN116083297A (en) | Microbial agent for preventing and treating strawberry root rot and preparation method thereof | |
| CN111378595A (en) | Burkholderia agricultural biocontrol strain Ba1 and application thereof | |
| CN116004419A (en) | Bacillus atrophaeus CY-2, microbial inoculum, preparation method and application thereof | |
| CN111979151B (en) | Biocontrol strain and application thereof | |
| CN111808785B (en) | Honghuadan endophytic Bacillus belgii and application thereof | |
| CN118325740A (en) | Trichoderma harzianum and application thereof in corn disease control |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20201009 |