CN106942276A - Bacillus subtilis BAB 1 prevents and treats the application of cucurbits powdery mildew - Google Patents
Bacillus subtilis BAB 1 prevents and treats the application of cucurbits powdery mildew Download PDFInfo
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Abstract
The invention discloses applications of the bacillus subtilis strain BAB 1 on preventing and treating cucurbits powdery mildew, belong to field of biological control.The invention also discloses application of the microbial bacterial agent containing BAB 1 on preventing and treating cucurbits powdery mildew.Bacillus subtilis BAB 1 of the present invention is high to the drug effect of cucurbits powdery mildew, and average preventive effect provides a kind of effective prevention and controls more than 85.0% for cucurbits powdery mildew；Secondly, the resistance to the action of a drug is not likely to produce using the preventing and treating cucurbits powdery mildews of bacillus subtilis BAB 1, lasting medicine is good；In addition, without problem of environmental pollution, preparing simply, using simple, low cost.
Description
Technical field
The invention belongs to field of biological control, and in particular to bacillus subtilis strain BAB-1 is in preventing and treating cucurbits powdery mildew
On application.
Background technology
Cucurbits powdery mildew is one of topmost leaf diseases in melon production, and the disease is main by two spore powdery mildews
(Erysiphe cichoracearumhe) and single softgel shell bacterium (Sphaerotheca fuliginea) two kinds of fungies cause, China
Based on single softgel shell bacterium (Sphaerotheca fuliginea).Cucurbits powdery mildew main harm cucumber, muskmelon, cucurbita pepo, south
The crop such as melon and balsam pear, especially cucumber and muskmelon, which are that northern China greenhouse is main, plants vegetables and fruit, the generation of its powdery mildew, seriously
Influence the yield and quality of cucumber and muskmelon；Generally because cucumber caused by powdery mildew or the muskmelon underproduction are about 10%, and it is popular
Time is up to 30% or so.The germ can infect melon plant from Seedling Stage to Adult plant, with the popular strong, incidence of disease
High, short incubation period and the characteristics of infect frequent again, is urgent problem to be solved in melon production.
Cucurbits powdery mildew is based on chemical prevention, such as thiophanate, triadimefon, auspicious malicious copper, carbendazim, Flusilazole, celestial life, generation
High is the primary chemical agricultural chemicals for preventing and treating powdery mildew of cucumber.But chemical pesticide is largely used for a long time, hold with pathogen
The resistance to the action of a drug is also easy to produce, causes prevention effect to reduce, while easily causing agricultural product and environmental pollution, causing new expression or secondary sick
The problems such as evil occurs greatly.Biological control is due to specialization is strong, lasting medicine is good, being not likely to produce the resistance to the action of a drug, do not result in
Environmental pollution and low cost and other advantages, are increasingly subject to favor.The microorganism of preventing and treating cucurbits powdery mildew mainly has at present：Lichens bud
Spore bacillus (Bacillusli cheniformis, CN102399712A), Trichoderma viride (Trichodermaviride,
CN101724572A), jute streptomycete (Streptomyces corchorusii, CN102870821A) bacillus subtilis
(Bacillus subtilis, CN1934241A, CN104403970A), Strepiomyces lavendulae (Strepomyces
Lavendulae, CN101481669A), trichoderma harzianum (Trichoderma harziamum, CN101575574A) and gill fungus shape
Bacillus (Bacillus mycoides, CN105002120A) etc., but the microorganism formulation of commercialization is simultaneously few.In addition,
It is many with the biology for tackling pathogen due to bio-diversity and common evolutionary effect, it is necessary to constantly screen new biological micro-organism
Sample and the continuous evolution of biological strain.
Patent《A kind of bacillus subtilis and its microbial inoculum and application》(CN101148650A) bacillus subtilis is disclosed in
Bacteria strain BAB-1 is in preventing and treating cotton rhizoctonia solani, rhizoctonia cerealis, botrytis cinerea, botrytis cinerea pers, tomato early epidemic
Germ, cucumber fusarium axysporum, stenocarpella maydis, fusarium graminearum, cotton-wilt fusarium, Maize yellow spot bacterium and wheat are complete
The application in the plant diseases such as germ is lost, but does not find the report by BAB-1 for preventing and treating cucurbits powdery mildew.
The content of the invention
Present invention aims at provide purposes of the bacillus subtilis strain BAB-1 on preventing and treating cucurbits powdery mildew.
Another object of the present invention is the purposes for providing the microbial bacterial agent containing BAB-1 on preventing and treating cucurbits powdery mildew.
3rd purpose of the invention is to provide the method that cucurbits powdery mildew is prevented and treated using bacillus subtilis strain BAB-1.
The present invention is achieved through the following technical solutions：
The invention provides bacillus subtilis (Bacillus subtilis) bacterial strain BAB-1 in preventing and treating cucurbits powdery mildew
On application.
Described bacillus subtilis (Baci11us subtilis) bacterial strain BAB-1 deposit number is CGMCC
No.2099 (referring to CN101148650A).
Cucurbits powdery mildew described in above-mentioned application refers to powdery mildew of cucumber or melon powdery mildew etc..
The pathogen of cucurbits powdery mildew described in above-mentioned application is two spore powdery mildews (Erysiphe
) or melon list softgel shell bacterium (Sphaerotheca fuliginea) cichoracearumhe.
Present invention also offers the answering on preventing and treating cucurbits powdery mildew of the microbial bacterial agent containing bacillus subtilis BAB-1
With.
The active component of described microbial bacterial agent is bacillus subtilis BAB-1 thalline.
Described microbial bacterial agent is liquid preparation.
The preparation method of described microbial bacterial agent, referring to CN101148650A specifications.
Described cucurbits powdery mildew refers to powdery mildew of cucumber or melon powdery mildew etc..
The pathogen of described cucurbits powdery mildew is two spore powdery mildews (Erysiphe cichoracearumhe) or melon list
Softgel shell bacterium (Sphaerotheca fuliginea).
Present invention also offers the side that cucurbits powdery mildew is prevented and treated using the microbial bacterial agent containing bacillus subtilis BAB-1
Method, including it is 10 that the microbial bacterial agent containing bacillus subtilis BAB-1 is diluted with water into viable bacteria body number7Cfu/mL, in melon
Class powdery mildew premorbid carries out foliar spray.
The present invention has the advantage that and beneficial effect：(1) present invention provides a kind of effective life for preventing and treating cucurbits powdery mildew
Thing prevention and controls；(2) bacillus subtilis BAB-1 is high to the drug effect of cucurbits powdery mildew, and average preventive effect is more than 85.0%；(3)
It is strong using bacillus subtilis BAB-1 and its microbial inoculum preventing and treating cucurbits powdery mildew specialization, and it is not likely to produce the resistance to the action of a drug；(4) this hair
The bright microbial bacterial agent being related to is to people, animal safety, without problem of environmental pollution；(5) prepared by microbial bacterial agent of the present invention
Method is simple, low cost, using simple.
Embodiment
The present invention is further clearly explained with specific embodiment below, but is not constituted in any way to the present invention
Limitation.Experimental method in following embodiments, unless otherwise instructed, is conventional method；Percentage in following embodiments contains
Amount, unless otherwise instructed, is weight percentage.
The preparation of the BAB-1 microbial bacterial agents of embodiment 1
Carry out in accordance with the following steps：
(1) actication of culture：- 80 DEG C bacillus subtilis (Baci11us subtilis) bacterial strain BAB-1 will be stored in
(deposit number is CGMCC No.2099) (its constituent and its weight ratio are in LB plating mediums：Tryptone 1g, ferment
Female extract 0.5g, sodium chloride 1g, agar powder 15g, water 1000mL) on activated (30 DEG C), picking single bacterium colony is on LB inclined-planes
Cultivated 20 hours at 32 DEG C on culture medium (its constituent and its weight ratio are with LB plating mediums), obtain the bacterial strain of activation；
(2) preparation of seed liquor：Making LB fluid nutrient mediums according to a conventional method, (its constituent and its weight ratio are：Pancreas
Peptone 1g, yeast extract 0.5g, sodium chloride 1g, water 1000mL), LB nutrient solution 100mL are loaded in 250mL triangular flasks,
High pressure moist heat sterilization, accesses in an oese step (1) bacterial strain activated after temperature drops to room temperature, in every bottle, 30 DEG C,
Shaken cultivation is carried out under conditions of shaking speed 190rpm 24h hours, obtain seed liquor；
(3) preparation of sucrose soya-bean cake culture medium：According to percentage by weight by sucrose 3.0%, beancake powder 2%, NaCl
0.1%, CaCO30.3%, KH2PO40.02% and MgSO4·7H2O 0.03% is added to the water, and is uniformly mixed, produces sugarcane
Sugared soya-bean cake culture medium；It is sub-packed in 500mL triangular flasks, every bottle of 200mL；Sterilizing 30 is carried out to sucrose soya-bean cake culture medium at 121 DEG C
Minute, then cool to 30 DEG C it is standby；
(4) fermented and cultured：The seed obtained by inoculation step (2) into every bottle of sucrose soya-bean cake culture medium 200mL obtained by step (3)
Liquid 2mL；Fermented and cultured 36h is carried out under the conditions of 30 DEG C, shaking speed 190rpm, was sampled later every 30 minutes from triangular flask
Microscopy is carried out, the gemma in the visual field and total thalline number are counted, and calculate gemma rate (gemma rate (%)=grown spore
Number/(grown spore number+thalline number) × 100)；Gemma rate stops fermented and cultured when reaching 90%；Common fermentation culture about 48h, is obtained
Bacillus subtilis BAB-1 liquid preparation.
The BAB-1 microbial bacterial agents of embodiment 2 are to the potted plant contrast test of powdery mildew of cucumber preventive effect
(1) test material：
(1) cucumber variety：Excellent No. 1 of Tianjin.
(2) powdery mildew of cucumber pathogen (Sphaerotheca fuliginea) SF-1 conidial suspensions.Initial bacterium
Strain comes from Agricultural University Of Hebei, and inoculation is with powdery mildew of cucumber pathogen conidial suspension by initial strains in Hebei province's agricultural
Numerous get is expanded in academy of sciences's Plant Protection Institute disease flocking biocontrol laboratory.
(2) test process：
(1) BAB-1 liquid preparations：100 times of water diluents of BAB-1 liquid preparations prepared by embodiment 1.
(2) blank control：Clear water.
(3) test method：
This experiment is carried out in Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie's disease flocking biocontrol laboratory.In diameter 6cm
Nutritive cube in cultivate excellent No. 1 cucumber seedling in Tianjin, seedling medium is vermiculite, and each nutritive cube sows two seeds, is left and taken after emerging
One strong sprout.Treat that cucumber seedling cotyledon flattens development experiment completely.Experiment sets 2 processing, and processing 1 is that reality is sprayed in cucumber cotyledons
100 times of water diluents of BAB-1 liquid preparations of the preparation of example 1 are applied, processing 2 is blank control, sprays equivalent clear water；Each processing weight
It is multiple 4 times, each repeat 20 plants of cucumber seedlings.The moisturizing 24h under the conditions of 25 DEG C, then spray inoculation powdery mildew of cucumber pathogen SF-1
Conidial suspension (1 × 105Individual/mL), in 25 DEG C incubated (illumination 14 hours, dark 10 hours).Treat blank control
After cotyledon is fully fallen ill, investigate the cucumber cotyledons powdery mildew state of an illness and calculate disease index and preventive effect.Investigation method, grade scale and
Prevention effect computational methods are with reference to National Standard of the People's Republic of China's pesticide field efficacy medicine test criterion (one) bactericidal agent for preventing and treating
Powdery mildew of cucumber (GB/T17980.30-2000).
As a result (being shown in Table 1), the cucumber through the processing of bacillus subtilis (Bacillus subtilis) BAB-1 liquid preparations is sub
The disease index of leaf is 1.37, the disease index 31.83 of substantially less than blank control, and BAB-1 liquid preparations are to powdery mildew of cucumber
Preventive effect be 95.70%.Illustrate that bacillus subtilis strain BAB-1 and its BAB-1 liquid preparation have to powdery mildew of cucumber prominent
The prevention effect gone out.
The BAB-1 liquid preparations of table 1 are to powdery mildew of cucumber preventive effect comparative test result
| Processing | Disease index | Preventive effect (%) |
| BAB-1 liquid preparations | 1.37b | 95.70 |
| Blank control | 31.83a | / |
The BAB-1 microbial bacterial agents of embodiment 3 are to the potted plant contrast test of powdery mildew of cucumber preventive effect
(1) test material：
(1) cucumber variety：Excellent No. 1 of Tianjin.
(2) powdery mildew of cucumber pathogen (Sphaerotheca fuliginea) SF-1 conidial suspensions are (with implementation
Example 2).
(2) test process：
(1) BAB-1 liquid preparations：100 times of water diluents of BAB-1 liquid preparations prepared by embodiment 1；
(2) blank control：Clear water.
(3) test method：
This experiment is carried out in Inst. of Plant Protection, Hebei-Prov. Academy of Agricultural and Forestry Scie's disease flocking biocontrol laboratory.In diameter 6cm
Nutritive cube in cultivate excellent No. 1 cucumber seedling in Tianjin, seedling medium is vermiculite, and each nutritive cube sows two seeds, is left and taken after emerging
One strong sprout.Be transplanted to when cucumber seedling grows two panels true leaf in diameter 15cm plastic flowerpot (in flowerpot load vermiculite with
Rural area soil is according to 1:The matrix that 1 ratio is made into), 25 DEG C incubated (illumination 14 hours, dark 10 hours).Cucumber seedling is used
Bamboo pole support is fixed, and carries out experiment after 5-7 piece true leaves are grown.Processing 1 sprays the BAB-1 liquid preparations 100 of the preparation of embodiment 1
Times water diluent；Processing 2 is blank control, sprays equivalent clear water.Each processing is repeated 4 times, each to repeat 5 basin cucumber seedlings.
Moisturizing 24h under the conditions of 25 DEG C；Then spray inoculation powdery mildew of cucumber pathogen SF-1 conidial suspensions (1 × 105Individual/
ML), continue at 25 DEG C incubated (illumination 14 hours, dark 10 hours).After blank control true leaf is fully fallen ill, investigation is yellow
The melon true leaf powdery mildew state of an illness simultaneously calculates disease index and preventive effect (GB/T17980.30-2000).
As a result the disease index for the cucumber true leaf for (being shown in Table 2) and being handled through BAB-1 liquid preparations is 6.42, substantially less than blank
The disease index 87.05 of control；BAB-1 liquid preparations are 92.62% to the preventive effect of powdery mildew of cucumber.Illustrate bacillus subtilis
Bacterial strain BAB-1 and its BAB-1 liquid preparation have prominent prevention effect to powdery mildew of cucumber.
The BAB-1 liquid preparations of table 2 are to powdery mildew of cucumber preventive effect comparative test result
| Processing | Disease index | Preventive effect (%) |
| BAB-1 liquid preparations | 6.42b | 92.62 |
| Blank control | 87.05a | / |
The BAB-1 microbial bacterial agents of embodiment 4 are to powdery mildew of cucumber preventive effect field comparison test
(1) test material：
Cucumber variety：Excellent No. 1 of Tianjin.
(2) test process：
(1) BAB-1 liquid preparations：100 times of water diluents of BAB-1 liquid preparations prepared by embodiment 1.
(2) chemical agent：10% difenoconazole water dispersible granule (Switzerland first just reaches crop protection Co., Ltd).
(3) blank control：Clear water.
(3) test method：
This, which is tested, satisfies in spring in 2015 in Hebei province Baoding Xushui County in cities and towns north of the city village village poplar Golconda cucumber booth
Carry out.Plough, apply fertilizer, sowing, watering with local farming operation.Experiment sets 3 processing, and processing 1 is microbial bacterial agent, is sprayed
100 times of water diluents of BAB-1 liquid preparations prepared by embodiment 1；Processing 2 is chemical agent, sprays 10% Difenoconazole water
800 times of water diluents of dispersible granule (Switzerland is first just producing up to crop protection Co., Ltd)；Processing 3 is blank control, is sprayed
Measure clear water；Each processing is repeated 4 times, each to repeat 3 rows, often about 25 cucumber seedlings of row.Start before powdery mildew of cucumber morbidity
Experiment, is spaced processing in 7-10 days once, until blank control is fully fallen ill, investigates the powdery mildew of cucumber state of an illness and calculates disease index
With preventive effect (GB/T17980.30-2000).
As a result (being shown in Table 3), the disease index through the processing of BAB-1 liquid preparations is 6.28, the substantially less than state of an illness of blank control
Index 76.25；BAB-1 liquid preparations to the preventive effect of powdery mildew of cucumber up to 91.76%, better than the prevention effect of chemical agent
89.04%.Illustrate that bacillus subtilis strain BAB-1 and its liquid preparation have prominent prevention effect to powdery mildew of cucumber.
The BAB-1 liquid preparations of table 3 are to powdery mildew of cucumber preventive effect comparative test result
| Processing | Disease index | Preventive effect (%) |
| BAB-1 liquid preparations | 6.28bc | 91.76 |
| Chemical agent | 8.36b | 89.04 |
| Blank control | 76.25a | / |
The BAB-1 microbial bacterial agents of embodiment 5 are to melon powdery mildew preventive effect field comparison test
(1) test material：
Melon variety：Radix curcumae is fragrant and red auspicious red, at Agricultural University Of Hebei's science and technology market seed sale.
(2) test process：
(1) BAB-1 liquid preparations：100 times of water diluents of BAB-1 liquid preparations prepared by embodiment 1.
(2) chemical agent：10% difenoconazole water dispersible granule (source be the same as Example 4).
(3) blank control：Clear water.
(3) test method：
This is tested carries out in spring in 2015 in Liu iron army of Zhang Dengdian villages of Hebei province Baoding Qingyuan County muskmelon greenhouse.Examination
Test and carry out simultaneously on two melon varieties.Experiment sets 3 processing, and processing 1 is microbial bacterial agent, sprays embodiment 1 and prepares
100 times of water diluents of BAB-1 liquid preparations；Processing 2 is chemical agent, sprays 10% difenoconazole water dispersible granule (auspicious
Scholar is first just producing up to crop protection Co., Ltd) 800 times of water diluents；Processing 3 is blank control, sprays equivalent clear water.Each
Processing is repeated 4 times, each to repeat 3 rows, often 20 muskmelon seedlings of row.Start experiment before melon powdery mildew morbidity, be spaced 7-10
Its processing once, until blank control is fully fallen ill, investigates the melon powdery mildew state of an illness and calculates disease index and preventive effect (reference
GB/T17980.30-2000)。
As a result (4 are shown in Table) in the fragrant field plot trial of melon variety radix curcumae, are handled through BAB-1 liquid preparations and chemistry
The disease index of chemicals treatment is respectively 11.1 and 13.0, the disease index (100.0) of substantially less than blank control, prevention effect
Respectively 88.9% and 87.0%；In the red auspicious red field plot trial of melon variety, handle and change through BAB-1 liquid preparations
The disease index for learning chemicals treatment is respectively 11.7 and 12.2, the disease index 100.0 of substantially less than blank control, prevention effect
Respectively 88.3% and 87.8%.Illustrate that there is BAB-1 and its BAB-1 liquid preparations preventing and treating well to imitate to melon powdery mildew
Really.
The BAB-1 liquid preparations of table 4 are to melon powdery mildew preventive effect comparative test result
Claims (9)
1. applications of bacillus subtilis (Bacillus subtilis) the bacterial strain BAB-1 on preventing and treating cucurbits powdery mildew.
2. application according to claim 1, it is characterised in that described cucurbits powdery mildew refers to powdery mildew of cucumber or muskmelon
Powdery mildew.
3. application according to claim 1, it is characterised in that the pathogen of described cucurbits powdery mildew is two spore powdery mildews
(Erysiphe cichoracearumhe) or melon list softgel shell bacterium (Sphaerotheca fuliginea).
4. application of the microbial bacterial agent containing bacillus subtilis BAB-1 on preventing and treating cucurbits powdery mildew.
5. application according to claim 4, it is characterised in that the active component of described microbial bacterial agent is withered grass gemma
Bacillus BAB-1 thalline.
6. thoroughly do away with the application described in claim 4, it is characterised in that described microbial bacterial agent is liquid preparation.
7. application according to claim 4, it is characterised in that described cucurbits powdery mildew refers to powdery mildew of cucumber or muskmelon
Powdery mildew.
8. application according to claim 4, it is characterised in that the pathogen of described cucurbits powdery mildew is two spore powdery mildews
(Erysiphe cichoracearumhe) or melon list softgel shell bacterium (Sphaerotheca fuliginea).
9. the method for cucurbits powdery mildew is prevented and treated using the microbial bacterial agent containing bacillus subtilis BAB-1, it is characterised in that will
It is 10 that microbial bacterial agent containing bacillus subtilis BAB-1, which is diluted with water to viable bacteria body number,7Cfu/mL, in cucurbits powdery mildew hair
Foliar spray is carried out before sick.
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| CN111763629A (en) * | 2019-03-27 | 2020-10-13 | 北京奥沃伽科技有限公司 | Bacillus belgii and application thereof |
| CN112400870A (en) * | 2020-11-27 | 2021-02-26 | 云南省微生物发酵工程研究中心有限公司 | Bacillus licheniformis water dispersible granule and application thereof |
| CN114806966A (en) * | 2022-05-20 | 2022-07-29 | 河北农业大学 | Pesticide-resistant Kulas-resistant wheat holoetch disease biological control bacillus subtilis and application thereof |
| CN115044491A (en) * | 2021-08-16 | 2022-09-13 | 金华市农业科学研究院(浙江省农业机械研究院) | Bacillus subtilis, microbial inoculum containing same, plant disease control agent and application of bacillus subtilis and microbial inoculum |
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