CN114196565B - Bacillus atrophaeus XZRKZ-23 and application thereof - Google Patents
Bacillus atrophaeus XZRKZ-23 and application thereof Download PDFInfo
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- CN114196565B CN114196565B CN202111197527.4A CN202111197527A CN114196565B CN 114196565 B CN114196565 B CN 114196565B CN 202111197527 A CN202111197527 A CN 202111197527A CN 114196565 B CN114196565 B CN 114196565B
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- 241000193410 Bacillus atrophaeus Species 0.000 title claims abstract description 48
- 235000020232 peanut Nutrition 0.000 claims abstract description 28
- 235000017060 Arachis glabrata Nutrition 0.000 claims abstract description 27
- 235000010777 Arachis hypogaea Nutrition 0.000 claims abstract description 27
- 235000018262 Arachis monticola Nutrition 0.000 claims abstract description 27
- 238000004321 preservation Methods 0.000 claims abstract description 3
- 241001558929 Sclerotium <basidiomycota> Species 0.000 claims description 12
- 241000894006 Bacteria Species 0.000 claims description 10
- 230000035784 germination Effects 0.000 claims description 10
- 230000002401 inhibitory effect Effects 0.000 claims description 5
- 241001530056 Athelia rolfsii Species 0.000 claims description 4
- 241001553178 Arachis glabrata Species 0.000 claims 1
- 244000105624 Arachis hypogaea Species 0.000 abstract description 28
- 230000000694 effects Effects 0.000 abstract description 10
- 240000001606 Adenanthera pavonina Species 0.000 abstract description 3
- 235000011470 Adenanthera pavonina Nutrition 0.000 abstract description 3
- 229930182764 Polyoxin Natural products 0.000 abstract description 3
- YEBIHIICWDDQOL-YBHNRIQQSA-N polyoxin Polymers O[C@@H]1[C@H](O)[C@@H](C(C=O)N)O[C@H]1N1C(=O)NC(=O)C(C(O)=O)=C1 YEBIHIICWDDQOL-YBHNRIQQSA-N 0.000 abstract description 3
- 230000001225 therapeutic effect Effects 0.000 abstract description 3
- 230000005764 inhibitory process Effects 0.000 description 15
- 241000196324 Embryophyta Species 0.000 description 13
- 201000010099 disease Diseases 0.000 description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 8
- 238000011282 treatment Methods 0.000 description 6
- 244000063299 Bacillus subtilis Species 0.000 description 5
- 235000014469 Bacillus subtilis Nutrition 0.000 description 5
- 230000000844 anti-bacterial effect Effects 0.000 description 5
- 230000001580 bacterial effect Effects 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 241000233866 Fungi Species 0.000 description 4
- 230000003042 antagnostic effect Effects 0.000 description 4
- 239000008223 sterile water Substances 0.000 description 4
- 208000024891 symptom Diseases 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 244000052616 bacterial pathogen Species 0.000 description 3
- 230000000443 biocontrol Effects 0.000 description 3
- 235000013339 cereals Nutrition 0.000 description 3
- 230000006378 damage Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 230000002262 irrigation Effects 0.000 description 3
- 238000003973 irrigation Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000002265 prevention Effects 0.000 description 3
- 230000009885 systemic effect Effects 0.000 description 3
- 229920000742 Cotton Polymers 0.000 description 2
- 244000299507 Gossypium hirsutum Species 0.000 description 2
- 206010039509 Scab Diseases 0.000 description 2
- 241000209140 Triticum Species 0.000 description 2
- 235000021307 Triticum Nutrition 0.000 description 2
- 239000013043 chemical agent Substances 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 241000192705 Aphanothece Species 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 241000195493 Cryptophyta Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 206010061217 Infestation Diseases 0.000 description 1
- 108010028921 Lipopeptides Proteins 0.000 description 1
- 235000007688 Lycopersicon esculentum Nutrition 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- 241000918585 Pythium aphanidermatum Species 0.000 description 1
- 241000221662 Sclerotinia Species 0.000 description 1
- 240000003768 Solanum lycopersicum Species 0.000 description 1
- 244000061456 Solanum tuberosum Species 0.000 description 1
- 235000002595 Solanum tuberosum Nutrition 0.000 description 1
- 241000082085 Verticillium <Phyllachorales> Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000007865 diluting Methods 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 235000012055 fruits and vegetables Nutrition 0.000 description 1
- 230000002538 fungal effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 150000002475 indoles Chemical class 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 235000021374 legumes Nutrition 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 244000005700 microbiome Species 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 244000052769 pathogen Species 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 235000012015 potatoes Nutrition 0.000 description 1
- 238000004382 potting Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 229930000044 secondary metabolite Natural products 0.000 description 1
- 238000009331 sowing Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000002255 vaccination Methods 0.000 description 1
Classifications
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Health & Medical Sciences (AREA)
- Plant Pathology (AREA)
- Microbiology (AREA)
- Pest Control & Pesticides (AREA)
- Biotechnology (AREA)
- Virology (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Environmental Sciences (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses a bacillus atrophaeus XZRKZ-23%Bacillus atrophaeus) The classification is named as bacillus atrophaeus @Bacillus atrophaeus) XZRKZ-23 was preserved in 2021 at 10 months and 12 days to China Center for Type Culture Collection (CCTCC) at the site of eight paths of Lopa nationality (Lopa nationality) in Wuchang district of Wuhan, hubei province, with the preservation number of CCTCC NO: m20211260. Bacillus atrophaeus XZRKZ-23 has good therapeutic effect on peanut southern blight, and is shown in the range of 10 6 At the dosage of/mL, the control effect is 21.1%, which is obviously higher than that of 200mg/L polyoxin (16.7%), showing that the bacillus atrophaeus XZRKZ-23 has great application prospect in preventing and treating peanut southern blight.
Description
Technical Field
The invention relates to bacillus atrophaeus.
Background
The peanut southern blight is a fungal soil-borne disease caused by sclerotium rolfsii (Sclerotium rolfsii sacc.) and has a wide host range, and can infect plants such as cereals, potatoes, legumes, fruits and vegetables. The peanut can be infected in the whole growing period, the basal part of the peanut stem is brown and soft in the later period of attack, the main stem, the side branches and the whole plant are yellow or wilted, and white hypha can be seen at the basal part of the stem. Under adverse conditions and at the late stage of disease, hyphae spread around the soil to form dark brown rapeseed-like sclerotium. After infestation of the pods, the hyphae cover the pods and the pods appear as brown rot in water. The peanut southern blight occurs in major peanut production countries in the world, the incidence rate in the field reaches 10% -40%, the yield loss can reach 20% -30%, and the serious disease can reach 80%. In China, henan, shandong, anhui, liaoning, jiangxi provinces and the like are reported. In recent years, the area is gradually enlarged, the yield and quality of peanuts are seriously influenced, and the method becomes an important factor for restricting the peanut production in China. In view of the damage to the Henan peanut industry, henan agricultural rural area parcels 2020 list peanut southern blight as Henan province class II pest and disease damage directory. Therefore, effective measures are taken to control the occurrence and harm of peanut southern blight, which is a problem to be solved urgently. At present, the prevention and control of peanut southern blight mainly depend on chemical agents. However, the use of chemical agents will inevitably have some negative effects, such as food safety of agricultural products, environmental pollution, drug resistance, etc.
Bacillus is an important class of microorganisms that are ubiquitous in vaccination in different environments. The aim of controlling plant diseases can be achieved by inhibiting or killing pathogens through competition, growth promotion, plant resistance induction, antagonistic substance direct generation and the like. Wherein, the bacillus atrophaeus (Bacillus atrophaeus) is a subspecies of the bacillus subtilis (Bacillus atrophaeus) and has a strong application prospect in the aspect of controlling plant diseases. The secondary metabolite, lipopeptides, indole compounds and the like of the bacillus atrophaeus have great application value in controlling plant fungal diseases, harmful algae and the like. Xin Haifeng it was found in the study that Bacillus atrophaeus reduced wheat scab and reduced wheat scab loss. Chen Li and the like have better control effects on cotton seedling damping off, cotton verticillium wilt and tomato root rot.
Disclosure of Invention
The invention aims to solve the technical problem of providing bacillus atrophaeus strain which has an inhibiting effect on the growth of hyphae of peanut southern blight bacteria.
The technical scheme of the invention is as follows: bacillus atrophaeus XZRKZ-23 (Bacillus atrophaeus), which is classified and named as Bacillus atrophaeus (Bacillus atrophaeus) XZRKZ-23, has been preserved in 2021 at 10 months and 12 days to China center for type culture collection, CCTCC for short, in the Wuchang district of Wuhan, hubei province, eight-path Lopa nations, and has a preservation number of CCTCC NO: m20211260.
The beneficial effects of the invention are as follows: the bacillus atrophaeus XZRKZ-23 has a strong inhibition effect on the growth of the hyphae of the southern blight fungus. The southern blight bacteria can not grow around the bacillus atrophaeus XZRKZ-23 to form an obvious antibacterial zone, and the antibacterial rate reaches 64.71 percent, which is equivalent to that of the control biocontrol strain bacillus subtilis.
The bacillus atrophaeus XZRKZ-23 can obviously inhibit germination of sclerotium of the southern blight fungus. At a Bacillus atrophaeus concentration of 10 3 The inhibition rate is more than 50 percent when the concentration is per mL, and the concentration is 10 5 When the inhibition rate reaches 81%, the concentration is 10 6 The inhibition rate is as high as 95.7% when the concentration is increased to 10% 7 In the above cases, germination of sclerotium of southern blight of peanut can be almost completely inhibited.
Bacillus atrophaeus XZRKZ-23 has good therapeutic effect on peanut southern blight, and is shown in the range of 10 6 At the dosage of/mL, the control effect is 21.1%, which is obviously higher than that of 200mg/L polyoxin (16.7%) of the control medicament, thus showing that the bacillus atrophaeus XZRKZ-23 has great application prospect in preventing and treating peanut southern blight.
Detailed Description
Bacillus atrophaeus is separated from peanut root soil and is used in preventing and controlling sclerotium rolfsii.
1. Test methods and results
1. Inhibition of bacillus atrophaeus XZRKZ-23 on growth of southern blight fungus hypha
The inhibition of the growth of the hyphae of the B.atrophaeus XZRKZ-23 was determined by the counter method. Firstly, a bacterial cake with the diameter of 5mm is made on the outer edge of an activated 2d peanut southern blight bacterial colony by a puncher, the bacterial cake is transferred to the center of a PDA flat plate, antagonistic bacillus atrophaeus XZRKZ-23 which is cultivated for 1d is inoculated at a position which is 2cm away from the edge of a culture dish by a streaking method, the bacterial cake is cultivated for 3d in dark at the temperature of 28 ℃, when the culture dish is full of contrast southern blight bacteria, the growth length of each treated mycelium is measured, the transverse mycelium length is measured for the treatment of inoculated antagonistic bacteria, and the bacteriostasis rate is calculated by adopting a formula (1). Commercial bacillus subtilis was isolated as a control strain. The test results are shown in Table 1.
TABLE 1 inhibition of B.atrophaeus XZRKZ-23 on hyphae of Sclerotinia arachidis (3 d)
| Testing biocontrol bacteria | Antibacterial rate |
| Bacillus atrophaeus XZRKZ-23 | 64.71％ |
| Bacillus subtilis | 61.45％ |
Table 1 shows that the bacillus atrophaeus XZRKZ-23 has a strong inhibition effect on the growth of the hyphae of the peanut southern blight bacteria. The southern blight bacteria can not grow around the bacillus atrophaeus XZRKZ-23 to form an obvious antibacterial zone, and the antibacterial rate reaches 64.71 percent, which is equivalent to that of the control biocontrol strain bacillus subtilis.
2. Inhibition of sclerotium germination of southern blight bacteria by bacillus atrophaeus XZRKZ-23
The activated bacillus atrophaeus XZRKZ-23 is transferred into 100mL LB liquid medium and shake cultured for 12h at 28 ℃ and 180 rpm. Then 1mL of the shaking-up fermentation broth was extracted, centrifuged at 5000rpm for 2min, and the supernatant was decanted. To the pellet, 1mL of sterile water was added, resuspended, and then diluted in a gradient to the following series of concentrations: 10 9 、10 8 、10 7 、10 6 、10 5 、 10 4 、10 3 And each mL. PDA medium was mixed with the bacterial suspension in a 9:1 ratio and then poured into 90mm plates. A mixture of sterile water and PDA medium was used as a control. 25 sclerotium were inoculated on each 90mm plate, three replicates were set for each treatment, and after 3d incubation at 28℃the number of sclerotium germination was counted. The germination of sclerotium was noted as the hyphal length exceeding 2mm on the horizontal plane. And calculating the sclerotium germination inhibition rate according to the formula (2). The test results are shown in Table 2.
TABLE 2 inhibition of sclerotium germination by B.atrophaeus XZRKZ-23 (3 d)
| Concentration (individual/mL) | 10 3 | 10 4 | 10 5 | 10 6 | 10 7 | 10 8 | 10 9 |
| Inhibition ratio (%) | 51.3 | 75.7 | 81 | 95.7 | 100 | 98.7 | 100 |
As can be seen from Table 2, the Bacillus atrophaeus XZRKZ-23 is capable of significantly inhibiting germination of sclerotium of Aphanothece arachidis hypogaeae. At a Bacillus atrophaeus concentration of 10 3 The inhibition rate is more than 50 percent when the concentration is per mL, and the concentration is 10 5 When the inhibition rate reaches 81%, the concentration is 10 6 The inhibition rate is as high as 95.7% when the concentration is increased to 10% 7 In the above cases, germination of sclerotium of southern blight of peanut can be almost completely inhibited.
3. Potted plant prevention and treatment effect of bacillus atrophaeus XZRKZ-23 on peanut southern blight
Selecting peanut seeds with full and consistent grain size, sowing the sterilized seeds into a flowerpot filled with a matrix in a greenhouse2 grains of each flowerpot, 7 days later, respectively inoculating the peanut southern blight germ, the bacillus atrophaeus XZRKZ-23 and a control medicament. Diluting the cultured fermentation broth with LB liquid medium until the thallus is 10 6 As a control agent, 200mg/L of the jinggangmycin solution was used per mL. Each peanut is inoculated with 2 southern fungus cakes with the diameter of 7mm, and then four treatments are set: (1) root irrigation of antagonistic bacteria fermentation liquor and southern blight pathogenic bacteria cake; (2) root irrigation with sterile water; (3) root irrigation with sterile water and southern blight and pathogenic bacteria cake; (4) the jinggangmycin irrigates roots and the southern pathogenic bacteria cakes. Each treatment was repeated 3 times, 10 strains each. The peanut plants were observed daily for onset and continued for 7 days after inoculation. Investigation was conducted according to the following disease classification criteria. Level 0: the plants are asymptomatic; stage 1: generating lesions only at the base of the stems; 2 stages: the basal part of the stem generates a constriction symptom, and less than 1/3 of the whole plant shows systemic symptoms (withering, death, wilting and the like); 3 stages: less than 2/3 of the whole plant shows systemic symptoms (wilting, death, wilting, etc.); 4 stages: more than 2/3 of the whole plant shows systemic symptoms (wilting, death, wilting, etc.); 5 stages: the whole plant dies. And (3) counting the disease index according to the formula (3), and calculating the relative prevention and treatment effect according to the formula (4). The potting test results are shown in Table 3.
TABLE 3 potted plant control effect of Bacillus atrophaeus XZRKZ-23 on peanut southern blight
As shown in Table 3, the bacillus atrophaeus XZRKZ-23 has good therapeutic effect on peanut southern blight, and the dose is 10 6 At the dosage of/mL, the control effect is 21.1 percent, which is obviously higher than that of 200mg/L polyoxin (16.7 percent) of the control medicament, which indicates that the bacillus atrophaeus XZRKZ-23 is used for preventing and controlling flowersHas great application prospect in the aspect of southern blight.
Claims (4)
1. Bacillus atrophaeus XZRKZ-23%Bacillus atrophaeus) The bacillus atrophaeus XZRKZ-23 is preserved in 2021 on 10-12 days to China center for type culture collection, and the preservation number is CCTCC NO: m20211260.
2. The use of bacillus atrophaeus XZRKZ-23 according to claim 1 for inhibiting the growth of hyphae of southern blight bacteria.
3. The use of bacillus atrophaeus XZRKZ-23 according to claim 1 for inhibiting sclerotium germination of sclerotium rolfsii.
4. The use of bacillus atrophaeus XZRKZ-23 according to claim 1 for controlling peanut southern blight.
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| CN114196565B true CN114196565B (en) | 2024-03-01 |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103281905A (en) * | 2010-12-21 | 2013-09-04 | 拜尔作物科学有限合伙公司 | Sandpaper mutants of bacillus and methods of their use to enhance plant growth, promote plant health and control diseases and pests |
| CN104531545A (en) * | 2014-09-11 | 2015-04-22 | 北京理工大学 | Bacillus atrophaeus strain DPPG-28 and application in crop disease control |
| CN106701623A (en) * | 2016-12-29 | 2017-05-24 | 山东农业大学 | Bacillus atrophaeus antagonistic to lycium chinensis root rot and application of bacillus atrophaeus |
| CN108641981A (en) * | 2018-05-17 | 2018-10-12 | 十堰市农业科学院 | One plant of Biocontrol Bacillus identification and application |
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- 2021-10-14 CN CN202111197527.4A patent/CN114196565B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103281905A (en) * | 2010-12-21 | 2013-09-04 | 拜尔作物科学有限合伙公司 | Sandpaper mutants of bacillus and methods of their use to enhance plant growth, promote plant health and control diseases and pests |
| CN104531545A (en) * | 2014-09-11 | 2015-04-22 | 北京理工大学 | Bacillus atrophaeus strain DPPG-28 and application in crop disease control |
| CN106701623A (en) * | 2016-12-29 | 2017-05-24 | 山东农业大学 | Bacillus atrophaeus antagonistic to lycium chinensis root rot and application of bacillus atrophaeus |
| CN108641981A (en) * | 2018-05-17 | 2018-10-12 | 十堰市农业科学院 | One plant of Biocontrol Bacillus identification and application |
Non-Patent Citations (1)
| Title |
|---|
| 暹罗芽孢杆菌ZHX-10的分离鉴定及其对花生白绢病的生防效果;张霞;许曼琳;郭志青;于静;吴菊香;于建垒;刘同金;李新国;迟玉成;万书波;;中国油料作物学报(第04期);全文 * |
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