CN103555617B - One bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof - Google Patents
One bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof Download PDFInfo
- Publication number
- CN103555617B CN103555617B CN201310494832.9A CN201310494832A CN103555617B CN 103555617 B CN103555617 B CN 103555617B CN 201310494832 A CN201310494832 A CN 201310494832A CN 103555617 B CN103555617 B CN 103555617B
- Authority
- CN
- China
- Prior art keywords
- microbial inoculum
- bacillus amyloliquefaciens
- disease
- fructus oleae
- oleae europaeae
- 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.)
- Expired - Fee Related
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof, the Classification And Nomenclature of this bacterial strain is bacillus amyloliquefaciens, Bacillus? amyloliquefaciens, complete called after bacillus amyloliquefaciens Ba4, being deposited in No. 1, North Star West Road, Chaoyang District, city of BeiJing, China institute No. 3 China Committee for Culture Collection of Microorganisms's common micro-organisms centers on September 12nd, 2013, does is deposit number CGMCC? No.8177.Bacillus amyloliquefaciens Ba4 provided by the invention can effective prevention and therapy Fructus oleae europaeae fruit decomposing disease.
Description
Technical field
The present invention relates to field of biology, in particular a bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof.
Background technology
Fructus oleae europaeae fruit decomposing disease (Alternariatenuissima) is Fructus oleae europaeae place of production kainogenesis disease, and mainly cause harm fruit, blade, cause shedding to fall leaves.Fruit susceptible rear be just black point, expand gradually, spot can be unified into large spot, and fruit abscission is rotten.There is no effective prevention and controls at present, particularly unpollution control methods technology does not relate to.Fructus oleae europaeae fruit is the important source material that sweet oil is produced, and therefore, adopts nuisanceless environment-friendly type measure to be the first-selection of preventing and treating this disease.
Therefore, prior art existing defects, needs to improve.
Summary of the invention
Technical problem to be solved by this invention is for the defect of prior art to Fructus oleae europaeae fruit preventing and treating rot disease, provides a bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof.
Technical scheme of the present invention is as follows:
One bacillus amyloliquefaciens, its Classification And Nomenclature is bacillus amyloliquefaciens, Bacillusamyloliquefaciens, be deposited in No. 1, North Star West Road, Chaoyang District, city of BeiJing, China institute No. 3 China Committee for Culture Collection of Microorganisms's common micro-organisms centers on 09 12nd, 2013, deposit number is CGMCCNo.8177.
The making method of the microbial inoculum of described bacillus amyloliquefaciens, be by nutrition meat nutrient solution after autoclaving, under logical oxygen controls, the bottling of 300mL triangular flask ratio is contained in 100mL liquid, inoculate bacillus amyloliquefaciens inclined-plane seed under aseptic conditions, every bottle graft kind 2 inclined-plane seeds, shaking culture, makes bacillus amyloliquefaciens microbial inoculum.
The making method of described microbial inoculum, the composition of its nutrition meat nutrient solution is extractum carnis 7.0g, peptone 10g, NaCl5g, water 1000mL.
The making method of described microbial inoculum obtains microbial inoculum.
The application of described microbial inoculum in prevention Fructus oleae europaeae fruit decomposing disease, is after the sick leaf of artificial removal, sick fruit, sprays described microbial inoculum spring, and every 10 days, every strain spray 200ml, continuous three times, can control disease and occur.
The application of described microbial inoculum in prevention Fructus oleae europaeae fruit decomposing disease, its bacteria suspension concentration is dilution less than 200 times.
The application of described microbial inoculum in treatment Fructus oleae europaeae fruit decomposing disease, sprays described microbial inoculum spring, and every 10 days, every strain spray 200ml, continuous three times, can significantly treat Fructus oleae europaeae fruit decomposing disease.
The application of described microbial inoculum in treatment Fructus oleae europaeae fruit decomposing disease, its bacteria suspension concentration is dilution less than 100 times.
Bacillus amyloliquefaciens Ba4 provided by the invention can effective prevention and therapy Fructus oleae europaeae fruit decomposing disease.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
1, the separation of endogenetic bacteria, purifying and preservation
Gather healthy Walnut Roots in walnut root rot spot, Chaotian District, Sichuan Guangyuan city, take back laboratory and be separated.The sample adopted back is clean with aseptic water washing, takes 1.0g root tissue, with the alcohol disinfecting 30s of 70%, after aseptic washing 3 times, then with after the NaClO sterilization 3min of 4%, aseptic washing 3 times, draw last washings 100 μ l and smear NA flat board (extractum carnis 3g, peptone 10g, sodium-chlor 5g, agar powder 15-20g, distilled water 1000ml, pH7.0,121 DEG C of autoclaving 30min after mixing packing), 28 DEG C of dark culturing 24h, for the contrast of surface sterilization, check that whether surface sterilization is thorough.
The root tissue of surface sterilization is shredded, puts into aseptic mortar, add quartz sand and the 10ml sterilized water of sterilizing, fully grind, leave standstill 30min, dilution 10
-1, 10
-2, 10
-3, 10
-4, 10
-5totally 5 gradients, draw 100 μ l10 respectively
-3, 10
-4, 10
-5these 3 gradient lapping liquids are coated with NA flat board (extractum carnis 3g respectively, peptone 10g, sodium-chlor 5g, agar powder 15-20g, distilled water 1000ml, pH7.0,121 DEG C of autoclaving 30min after mixing packing), each process repeats for 3 times, 28 DEG C of dark culturing 48h, choose the most uniform gradient of growth, carry out plate count.
According to the size of bacterium colony, color, whether projection, edge feature, smooth surface whether, the difference of the aspect such as transparency, picking list bacterium colony is at the flat lining out purifying of NA, and NA inclined-plane of transferring after purifying is for subsequent use in 4 DEG C of preservations.Obtain 10 endogenetic bacterias altogether.
2, endogenetic bacteria is to the dull and stereotyped antagonistic action of Fructus oleae europaeae fruit decomposing disease germ
Dull and stereotyped face-off method: at Fructus oleae europaeae fruit decomposing disease germ (Alternariatenuissima) the bacterium cake (diameter 5mm) of culture dish central authorities inoculation 7d, 3cm place is about from this germ cake, the endophyte cultivating 2d is picked respectively with transfering loop, at each standardized bar fine rule in the symmetrical both sides of germ, 28 DEG C of cultivations, only to inoculate the flat board of germ for contrast, in time contrasting and cover with ware, measure the colony growth diameter of germ, often process repetition 3 times.Be calculated as follows colony growth inhibiting rate:
Colony growth inhibiting rate=(contrast bacterium colony clean growth diameter-clean growth diameter of process bacterium colony)/contrast bacterium colony grows only
Diameter × 100%
Table 1 is that after 7d, endogenetic bacteria is to the antagonistic action of Fructus oleae europaeae fruit decomposing disease germ (Alternariatenuissima), and result shows, and have 5 kinds of endogenetic bacteria colony growth inhibiting rates to reach more than 75.39%, wherein Ba4 bacterial strain inhibiting rate reaches 98.46%.
Table 1 endogenetic bacteria is to the antagonistic action (7 days) of Fructus oleae europaeae fruit decomposing disease germ
Note: * otherness compares: be that difference is remarkable containing same letter, otherwise be remarkable.
3, the Identification of Species of interior raw Antagonistic Fungi
Through morphological observation, be B.amyloliquefaciens(bacillus amyloliquefaciens in conjunction with physiological and biochemical index (table 2) and this endophyte of molecular biology identification Ba4).
NA plate culture medium has been cultivated single bacteria colony white of the Ba4 bacterial strain of 2d, and rounded, edge is irregular, and surface drying has fold, inviscid, opaque, G
+.
Table 2 bacterial strain Ba4 physiological and biochemical index
4, greenhouse test
(1) Ba4 biocontrol microorganisms prepares: nutrition meat nutrient solution (extractum carnis 7.0g, peptone 10g, NaCl5g, water 1000mL) after autoclaving, be contained in 300mL triangular flask ratio (logical oxygen controls) bottling in 100mL liquid, at sterile state inoculation Ba4 inclined-plane seed, every bottle graft kind 2 inclined-plane seeds, temperature 26-28 DEG C, initial pH value 7.0,120r/min shaking culture 36h, makes Ba4 microbial inoculum.
(2) cause of disease inoculation and process
Choosing 4 years oil generation olives transplants in greenhouse, PDA inclined-plane olea europaea fruit maize ear rot bacterium (Alterniasp) conidium (concentration 3 × 10
5) inoculate fruit after one week, adopt spray method to divide different concns to carry out sprinkling process to Fructus oleae europaeae, every basin 200mL with (1) described microbial inoculum, and compare with sterilized water, repeat 3 times, statistics Fructus oleae europaeae incidence.Often repeat Fructus oleae europaeae 50 strain.
After the dilution of Ba4 fermentation liquor, show different prevention effect (table 3), Be very effective below stoste to 400 times, can substantially control disease and not occur by special less than 200 times.Based on this tests, carry out field test.
Prevention effect after table 3 greenhouse test 15d
5, field control example
5.1, prevent
The early spring selects the slight milpa of morbidity (sickness rate 15%, disease index about 5), manually except after leaf of preventing or cure a disease (really), spray the Ba4 microbial inoculum of different concns described in 4 spring, every 10 days, every strain spray 200ml, continuous three times, compared with clear water and adds up incidence summer.Less than 200 times basic controlling disease can there is (table 4).
Table 4 field preventive effect (control in the early spring, summer are added up)
Note: letter is for p<0.05 level has significant difference.
5.2, treat
Select olea europaea fruit maize ear rot over the years that serious milpa (sickness rate 100%, disease-sensitive index 64) occurs.Spray different concns Ba4 microbial inoculum described in 4 spring, every strain spray 200ml every 10 days, continuous three times, compare with clear water, summer adds up incidence, and table 5 illustrates that this bacteria preparation has remarkable therapeutic action, significantly reduces disease-sensitive index, and less than 100 times effects are best.Sickness rate and disease-sensitive index (table 6) are by following criterion calculation.
Table 5 Field treatment effect (control in spring, summer are added up)
Note: letter is for p<0.05 level has significant difference.
The grade scale of table 6 leaf, fruit disease evil
Should be understood that, for those of ordinary skills, can be improved according to the above description or convert, and all these improve and convert the protection domain that all should belong to claims of the present invention.
Claims (7)
1. the making method of the microbial inoculum of a bacillus amyloliquefaciens, it is characterized in that, by nutrition meat nutrient solution after autoclaving, under logical oxygen controls, be contained in the bottling of 300mL triangular flask ratio in 100mL liquid, inoculate bacillus amyloliquefaciens inclined-plane seed under aseptic conditions, every bottle graft kind 2 inclined-plane seeds, shaking culture, makes bacillus amyloliquefaciens microbial inoculum; Described bacillus amyloliquefaciens deposit number is CGMCCNo.8177.
2. the making method of microbial inoculum according to claim 1, is characterized in that, the composition of nutrition meat nutrient solution is extractum carnis 7.0g, peptone 10g, NaCl5g, water 1000mL.
3. the making method of microbial inoculum according to claim 1 and 2 obtains microbial inoculum.
4. the application of microbial inoculum according to claim 3 in prevention Fructus oleae europaeae fruit decomposing disease, is characterized in that, after the sick leaf of artificial removal, sick fruit, sprays described microbial inoculum spring, and every 10 days, every strain spray 200ml, continuous three times, can control disease and occur.
5. the application of microbial inoculum according to claim 4 in prevention Fructus oleae europaeae fruit decomposing disease, it is characterized in that, bacteria suspension concentration is dilution less than 200 times.
6. the application of microbial inoculum according to claim 3 in treatment Fructus oleae europaeae fruit decomposing disease, is characterized in that, spray described microbial inoculum spring, and every 10 days, every strain spray 200ml, continuous three times, can significantly treat Fructus oleae europaeae fruit decomposing disease.
7. the application of microbial inoculum according to claim 6 in treatment Fructus oleae europaeae fruit decomposing disease, it is characterized in that, bacteria suspension concentration is dilution less than 100 times.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310494832.9A CN103555617B (en) | 2013-10-21 | 2013-10-21 | One bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310494832.9A CN103555617B (en) | 2013-10-21 | 2013-10-21 | One bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103555617A CN103555617A (en) | 2014-02-05 |
| CN103555617B true CN103555617B (en) | 2016-04-06 |
Family
ID=50009983
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310494832.9A Expired - Fee Related CN103555617B (en) | 2013-10-21 | 2013-10-21 | One bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103555617B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109897795A (en) * | 2018-12-26 | 2019-06-18 | 佛山市艳晖生物科技有限公司 | A kind of microbial bacterial agent and its application on anti-treat constipation |
| CN112322521B (en) * | 2020-10-23 | 2022-05-03 | 贵州大学 | Bacillus amyloliquefaciens GUOR0918 and application thereof in preventing and treating soft rot of kiwi fruits |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102839142A (en) * | 2012-09-14 | 2012-12-26 | 杨凌绿都生物科技有限公司 | Bacillus amyloliquefaciens (Ba 168) and fermenting culture method and application of bacillus amyloliquefaciens |
| CN102864100A (en) * | 2012-08-16 | 2013-01-09 | 北京雷力农用化学有限公司 | Bacillus amyloliquefaciens and application thereof |
-
2013
- 2013-10-21 CN CN201310494832.9A patent/CN103555617B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102864100A (en) * | 2012-08-16 | 2013-01-09 | 北京雷力农用化学有限公司 | Bacillus amyloliquefaciens and application thereof |
| CN102839142A (en) * | 2012-09-14 | 2012-12-26 | 杨凌绿都生物科技有限公司 | Bacillus amyloliquefaciens (Ba 168) and fermenting culture method and application of bacillus amyloliquefaciens |
Non-Patent Citations (1)
| Title |
|---|
| 解淀粉芽孢杆菌NK10.BAhjaWT抑真菌作用的研究;郝建安等;《微生物学通报》;20080620;第35卷(第6期);903-908 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103555617A (en) | 2014-02-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111172080A (en) | Bacillus belgii and application thereof | |
| CN109593681A (en) | A kind of composite bacteria agent and its preparation method and application for preventing and treating aquaculture cyanobacterial bloom | |
| CN109207404A (en) | Siam bacillus YJ15 and its application | |
| CN102703342B (en) | Bacillus velezensis ZJ20 strain and liquid preparations thereof | |
| CN103981102A (en) | DSE (Dark septate endophyte) bacterial strain 24L-4 and application thereof in dendrobium officinale production | |
| CN103224904A (en) | Rhodopseudomonas strain, biocontrol microbial inoculum and biocontrol fermentation liquid, and corresponding preparation methods and application thereof in controlling phytophthora blight of pepper | |
| CN103396960B (en) | Bacillus cereus (strain B2), liquid preparation, preparation method of liquid preparation, and application of B2 strain or liquid preparation in prevention and treatment of melanconium juglandinum kunze | |
| CN103555616B (en) | One bacillus amyloliquefaciens and the application of liquid preparation in control walnut root rot thereof | |
| CN108913622B (en) | Bacillus megaterium BM22 and preparation and application of spore powder thereof | |
| CN110317747A (en) | A kind of bacillus amyloliquefaciens JT68 and its application in prevention and treatment tea anthracnose | |
| CN103555618B (en) | Bacillus amyloliquefaciens and application of liquid preparations thereof in treatment of cenangium ferruginosum | |
| CN103555617B (en) | One bacillus amyloliquefaciens and the application of microbial inoculum in control Fructus oleae europaeae fruit decomposing disease thereof | |
| CN109321500A (en) | One bacillus amyloliquefaciens bacterial strain and its application in prevention and treatment Oil Tea Anthracnose evil | |
| CN108795827A (en) | A kind of bacillus megaterium BM22 and its water dispersible granules and application | |
| CN105018393A (en) | Bacillus megaterium strain and application thereof | |
| CN105462882B (en) | A kind of pseudomonas aeruginosa and its application for preventing crop verticillium wilt | |
| CN104673701B (en) | The preparation and application of one plant of phloridzin degradation bacteria and its microbial inoculum | |
| CN116676234A (en) | Salt-tolerant bacillus JK-25, microbial inoculum, preparation method and application thereof | |
| CN116536207A (en) | Bacillus atrophaeus WLKYSY-4, biological microbial inoculum and application thereof | |
| CN111471627B (en) | Serratia marcescens BSZ and application thereof | |
| CN102367429B (en) | Bacillus subtilis for controlling tobacco anthracnose | |
| CN105018395A (en) | Bacillus pumilus strain and application thereof in apple alternaria leaf spot prevention and control | |
| CN112741113B (en) | Microbial preparation for preventing and treating potato early blight and application thereof | |
| CN104845909B (en) | One plant of Gorgon fruit rhizosphere growth-promoting bacterium and its application and microbial inoculum | |
| CN105176895B (en) | A kind of bacillus amyloliquefaciens and its application for preventing cotton verticillium wilt |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20160406 Termination date: 20181021 |