CN102870821A - Application of metabolins of streptomyces corchorusii NF0919 bacterial strain in prevention and control cucumber diseases - Google Patents

Application of metabolins of streptomyces corchorusii NF0919 bacterial strain in prevention and control cucumber diseases Download PDF

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CN102870821A
CN102870821A CN2012103546314A CN201210354631A CN102870821A CN 102870821 A CN102870821 A CN 102870821A CN 2012103546314 A CN2012103546314 A CN 2012103546314A CN 201210354631 A CN201210354631 A CN 201210354631A CN 102870821 A CN102870821 A CN 102870821A
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fermentation
bacterial strain
seed culture
cucumber
hpo
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杨敬辉
陈宏州
张文文
庄义庆
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Zhenjiang Institute of Agricultural Sciences Jiangsu Hilly Area
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Zhenjiang Institute of Agricultural Sciences Jiangsu Hilly Area
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Abstract

The invention relates to a method for extracting active metabolins in fermentation broth of streptomyces corchorusii NF0919 bacterial strain (whose preservation number is CGMCC (China General Microbiology Culture Center) No. 3968 and strain patent number is ZL201010236886.1). The metabolins are capable of effectively preventing and controlling Erysiphecichoracearum, Pseudoperonosporacubensis, Botrytiscinerea and Fusariumoxysporum.

Description

The application of jute streptomycete NF0919 Metabolite on the control cucumber disease
Invention field
The present invention relates to a kind of microorganism and purposes, more specifically to a kind of jute streptomycete ( Streptomyces corchorusii) NF0919 Metabolite and the application in the control of control cucumber disease thereof, belong to biological pesticide technical field.
Background technology
Powdery mildew of cucumber ( Erysiphe cichoracearum), cucumber downy mildew ( Pseudoperonospora cubensis), gray mold of cucumber ( Botrytis cinerea) and cucumber fusarium axysporum ( Fusarium oxysporum) be fungi and oomycetes disease main on the cucumber.The control of cucumber disease is current mainly to come prevention and control by chemical control, and chemical agent commonly used has: triazolone, Sukeling fumicants, Wakil, Fluoxastrobin, kresoxim-methyl, Difenoconazole, Enestroburin, Boscalid and pyraclostrobin etc.For a long time, because multifrequency is inferior, multiple dose is used chemical agent, cause the cucumber pathogen that chemical agent commonly used has all been produced anti-(anti-) property of medicine in various degree, occur repeatedly the failed example of control in the production.
The biopesticide special preparation that successfully is used at present the control cucumber disease is less.Document reported the biological and ecological methods to prevent plant disease, pests, and erosion Pseudomonas (kind) of biological control control disease mainly contain the trichoderma fungi ( TrichodermaTrichoderma viride spp.) ( Trichoderma viride) and Trichoderma harzianum ( Trichoderma harzianum); Bacillus ( BacillusBacillus subtilis spp.) ( Bacillus subtilis), bacillus polymyxa ( Bacillus polymyxa) and bacillus megaterium ( Bacillus megaterium); The pseudomonas bacterium ( PseudomonasThe false single-cell bacteria of fluorescence spp.) ( Pseudomonas fluorescens) etc.Agricultural antibiotic mainly contains pesticide corrosion 120, kasugarnycin and Wuyiencin etc.
The bacterium jute streptomycete NF0919 bacterial strain of flying up and down of simultaneously double harness the Yellow River melon fungi and oomycetes disease provided by the invention is not mentioned in the prior art and is disclosed.
Summary of the invention
Provide the jute streptomycete ( Streptomyces corchorusii) extracting method of metabolite in the NF0919 bacterial strain fermentation liquor, and extract is in the application that prevents and treats on powdery mildew of cucumber, cucumber downy mildew, gray mold of cucumber and the cucumber fusarium axysporum.
The present invention is achieved by the following technical solutions:
The strain characteristics of the jute streptomycete NF0919 bacterial strain of usefulness is provided among the present invention, sees patent ZL 201010236886.1.Jute streptomycete (Latin name is Streptomyces corchorusii) NF0919 bacterial strain, in on July 8th, 2010 in China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation, the depositary institution address is No. 3 Institute of Microorganism, Academia Sinica in Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and deposit number is: CGMCC No. 3968.The extracting method of metabolite in the jute streptomycete NF0919 bacterial strain fermentation liquor of the present invention may further comprise the steps:
(A) cultivation of the activation of bacterial strain and first order seed culture fluid
The inclined-plane of the NF0919 bacterial strain preserved is activated at the Gause I medium, cultivate 10 d, picking 1 cm for 30 ℃ 2The bacterium piece of size is inoculated in (bottled culture fluid 50 mL of 250 mL triangles) in the 50 mL first order seed culture fluids, place 30 ℃ of shaken cultivation 44～48 h on the 170 rpm shaking tables, the composition of wherein said 50 mL first order seed culture fluids is: soluble starch 3%, glucose 2%, dusty yeast 1%, peptone 1%, NaCl 0.5%, K 2HPO 40.2%, soluble starch, glucose, dusty yeast, peptone, NaCl and K 2HPO 4The unit ratio of content is W/V;
(B) cultivation of secondary seed culture fluid
With step (A) gained first order seed culture fluid by 5%(V/V) inoculum concentration access (bottled culture fluid 50 mL of 250 mL triangles) in the 50 mL secondary seed culture fluids, cultivate 48-52 h on 170 rpm, 30 ℃ of shaking tables, wherein said 5% unit ratio is V/V; The composition (W/V) of described 50 mL secondary seed culture fluids: potato starch 8.5%, cottonseed protein 2.2%, CaCO 30.1%, K 2HPO 40.05%, MgSO 40.05%, amylase 0.01%, potato starch, cottonseed protein, CaCO 3, K 2HPO 4, MgSO 4With the unit ratio of amylase content be W/V;
(C) cultivation of fermentation culture
With step (B) gained secondary seed culture fluid by 5%(V/V) inoculum concentration to be inoculated in the volume that 160 L fermentation cultures are housed be in the fermentation tank of 200 L, wherein said 5% unit ratio is V/V, fermentation parameter is set is: dissolved oxygen 100%, and defoamer 1.6 L, mixing speed is 360 rpm, fermentation temperature is 36 ℃, fermentation time is 72 h, and pH 6.8～7.2, and the composition of described fermentation culture is (W/V): potato starch 8.5%, cottonseed protein 2.6%, K 2HPO 40.05%, NaCl 0.2%, MgSO 40.05%, CaCO 30.1%, pH7.2-7.4, potato starch, cottonseed protein, K 2HPO 4, NaCl, MgSO 4And CaCO 3The unit ratio of content is W/V; Described defoamer is XP-100F series fermentation silicone defoaming agent;
(D) metabolite separating method in the NF0919 bacterial strain fermentation liquor
Step (C) gained zymotic fluid is got supernatant at centrifugal 5 min of 10000 rpm, regulate pH to 2.0 with concentrated hydrochloric acid, precipitation occurs, again centrifugal, the collecting precipitation thing is used ethyl acetate extraction 3 times, and 35 ℃ of vacuum dryings get extract.
The invention has the beneficial effects as follows:
The invention provides the extracting method of metabolite in the jute streptomycete NF0919 bacterial strain fermentation liquor, and prove that its extract can prevent and treat the cucumber multiple diseases simultaneously.Control in field is the result show, this Metabolite all has good control efficiency to powdery mildew of cucumber, cucumber downy mildew, gray mold of cucumber and cucumber fusarium axysporum.
Embodiment
Embodiment
The strain characteristics of the jute streptomycete NF0919 bacterial strain of usefulness is provided among the present invention, sees patent ZL201010236886.1.The extracting method of product in the jute streptomycete NF0919 bacterial strain fermentation liquor of the present invention may further comprise the steps:
(A) cultivation of the activation of bacterial strain and first order seed culture fluid
The inclined-plane of the NF0919 bacterial strain preserved is activated at the Gause I medium, cultivate 10 d, picking 1 cm for 30 ℃ 2The bacterium piece of size is inoculated in (bottled culture fluid 50 mL of 250 mL triangles) in the 50 mL first order seed culture fluids, place 30 ℃ of shaken cultivation 44～48 h on the 170 rpm shaking tables, the composition of wherein said 50 mL first order seed culture fluids is: soluble starch 3%, glucose 2%, dusty yeast 1%, peptone 1%, NaCl 0.5%, K 2HPO 40.2%, soluble starch, glucose, dusty yeast, peptone, NaCl and K 2HPO 4The unit ratio of content is W/V;
(B) cultivation of secondary seed culture fluid
With step (A) gained first order seed culture fluid by 5%(V/V) inoculum concentration access (bottled culture fluid 50 mL of 250 mL triangles) in the 50 mL secondary seed culture fluids, cultivate 48-52 h on 170 rpm, 30 ℃ of shaking tables, wherein said 5% unit ratio is V/V; The composition (W/V) of described 50 mL secondary seed culture fluids: potato starch 8.5%, cottonseed protein 2.2%, CaCO 30.1%, K 2HPO 40.05%, MgSO 40.05%, amylase 0.01%, potato starch, cottonseed protein, CaCO 3, K 2HPO 4, MgSO 4With the unit ratio of amylase content be W/V;
(C) cultivation of fermentation culture
With step (B) gained secondary seed culture fluid by 5%(V/V) inoculum concentration to be inoculated in the volume that 160 L fermentation cultures are housed be in the fermentation tank of 200 L, wherein said 5% unit ratio is V/V, fermentation parameter is set is: dissolved oxygen 100%, and defoamer 1.6 L, mixing speed is 360 rpm, fermentation temperature is 36 ℃, fermentation time is 72 h, and pH 6.8～7.2, and the composition of described fermentation culture is (W/V): potato starch 8.5%, cottonseed protein 2.6%, K 2HPO 40.05%, NaCl 0.2%, MgSO 40.05%, CaCO 30.1%, pH7.2-7.4, potato starch, cottonseed protein, K 2HPO 4, NaCl, MgSO 4And CaCO 3The unit ratio of content is W/V; Described defoamer is XP-100F series fermentation silicone defoaming agent;
(D) extracting method of metabolite in the NF0919 bacterial strain fermentation liquor
Step (C) gained zymotic fluid is got supernatant at centrifugal 5 min of 10000 rpm, regulate pH to 2.0 with concentrated hydrochloric acid, precipitation occurs, again centrifugal, the collecting precipitation thing is used ethyl acetate extraction 3 times, and 35 ℃ of vacuum dryings get extract.
The implementation result of present embodiment:
The extract of NF0919 bacterial strain fermentation liquor is to the field efficiency test of powdery mildew of cucumber, cucumber downy mildew, gray mold of cucumber and cucumber fusarium axysporum.Experiment is at white rabbit town, Jurong City, Jiangsu Province green house, and cucumber variety is excellent No. two of Tianjin.The test processing scheme is as follows:
(A) control of powdery mildew of cucumber
Medicament processing method: 3000 times of liquid of NF0919 strain fermentation liquid extract, the contrast medicament is 3000 times of liquid of 50% water dispersion granules agent of Kresoxim (Jiangsu Province Lvdunzhibao Agriculture Chemical Experiment Co., Ltd sale), blank is not for preventing and treating residential quarter area 40 m 2, district's group is arranged at random, 4 repetitions.Processing time is in April, 2011, control interval 10 days, and thin spraying control is 3 times continuously, and water consumption is 50kg/667m 2The front investigation of for the first time dispenser also calculates state of an illness radix, prevents and treats for the last time rear 10 days investigation disease indexs;
(B) control of cucumber downy mildew
Medicament processing method: 3000 times of liquid of NF0919 strain fermentation liquid extract, the contrast medicament is 3000 times of liquid of 50% water dispersion granules agent of Kresoxim (Jiangsu Province Lvdunzhibao Agriculture Chemical Experiment Co., Ltd sale), blank is not for preventing and treating residential quarter area 40 m 2, district's group is arranged at random, 4 repetitions.Processing time is in May, 2011, control interval 10 days, and thin spraying control is 3 times continuously, and water consumption is 50kg/667m 2The front investigation of for the first time dispenser also calculates state of an illness radix, prevents and treats for the last time rear 10 days investigation disease indexs;
(C) control of gray mold of cucumber
Medicament processing method: 3000 times of liquid of NF0919 strain fermentation liquid extract, the contrast medicament is 3000 times of liquid of 50% water dispersion granules agent of Kresoxim (Jiangsu Province Lvdunzhibao Agriculture Chemical Experiment Co., Ltd sale), blank is not for preventing and treating residential quarter area 40 m 2, district's group is arranged at random, 4 repetitions.Processing time is in April, 2011, control interval 10 days, and thin spraying control is 3 times continuously, and water consumption is 50kg/667m 2The front investigation of for the first time dispenser also calculates state of an illness radix, prevents and treats for the last time rear 10 days investigation disease indexs;
(D) control of cucumber fusarium axysporum
Medicament processing method: 3000 times of liquid of NF0919 strain fermentation liquid extract, the contrast medicament is 3000 times of liquid of 50% water dispersion granules agent of Kresoxim (Jiangsu Province Lvdunzhibao Agriculture Chemical Experiment Co., Ltd sale), when transplanting, cucumber transplants cucumber seedling filling 200ml liquid for every, blank is filled with clear water, 40 every residential quarters of seedling, district's group is arranged at random, 4 repetitions.Fill with the root time in February, 2011,60 days " Invest, Then Investigate " disease indexs.
The field control result: the control efficiency of NF0919 strain fermentation liquid extract 3000 times of liquid powderys mildew of cucumber, cucumber downy mildew, gray mold of cucumber and cucumber fusarium axysporums be respectively 97.18%, 96.68%, 92.00% and 80.23%(table 1), and contrast medicament 50% water dispersion granules agent of Kresoxim 3000 times of liquid to the control efficiency of powdery mildew of cucumber, cucumber downy mildew, gray mold of cucumber and cucumber fusarium axysporum be respectively 90.63%, 86.29%, 73.15% and 33.07%(table 1).
Above result shows that the field control effect of metabolite extract is significantly higher than the contrast medicament in the NF0919 bacterial strain fermentation liquor, and NF0919 strain fermentation liquid extract is controlled effect significantly to the double of cucumber multiple diseases.The poisoning phenomenon does not all occur in all processing in the test, shows that metabolite uses safety in the NF0919 bacterial strain fermentation liquor.
The extract of table 1 NF0919 bacterial strain fermentation liquor is to the field controling test result of cucumber disease
Figure 2012103546314100002DEST_PATH_IMAGE001
Annotate: different capitalizations represent P＜0.01.

Claims (3)

  1. The jute streptomycete ( Streptomyces corchorusii) extracting method of metabolite in NF0919 bacterial strain (this strain properties the is seen patent ZL 201010236886.1) zymotic fluid.
  2. 2. the extract of metabolite in the jute streptomycete NF0919 bacterial strain fermentation liquor according to claim 1 is in the application that prevents and treats on powdery mildew of cucumber, cucumber downy mildew, gray mold of cucumber and the cucumber fusarium axysporum.
  3. 3. the extracting method of metabolite in the jute streptomycete NF0919 bacterial strain fermentation liquor claimed in claim 1 is characterized in that may further comprise the steps:
    (A) cultivation of the activation of bacterial strain and first order seed culture fluid
    The inclined-plane of the NF0919 bacterial strain preserved is activated at the Gause I medium, cultivate 10 d, picking 1 cm for 30 ℃ 2The bacterium piece of size is inoculated in the 50 mL first order seed culture fluids, places 30 ℃ of shaken cultivation 44～48 h on the 170 rpm shaking tables, and the composition of wherein said 50 mL first order seed culture fluids is: soluble starch 3%, glucose 2%, dusty yeast 1%, peptone 1%, NaCl 0.5%, K 2HPO 40.2%, soluble starch, glucose, dusty yeast, peptone, NaCl and K 2HPO 4The unit ratio of content is W/V;
    (B) cultivation of secondary seed culture fluid
    Step (A) gained first order seed culture fluid is accessed in the 50 mL secondary seed culture fluids by 5% inoculum concentration, cultivate 48-52 h on 170 rpm, 30 ℃ of shaking tables, wherein said 5% unit ratio is V/V; The composition of described 50 mL secondary seed culture fluids: potato starch 8.5%, cottonseed protein 2.2%, CaCO 30.1%, K 2HPO 40.05%, MgSO 40.05%, amylase 0.01%, potato starch, cottonseed protein, CaCO 3, K 2HPO 4, MgSO 4With the unit ratio of amylase content be W/V;
    (C) cultivation of fermentation culture
    It is in the fermentation tank of 200 L that step (B) gained secondary seed culture fluid is inoculated in the volume that 160 L fermentation cultures are housed by 5% inoculum concentration, wherein said 5% unit ratio is V/V, fermentation parameter is set is: dissolved oxygen 100%, defoamer (XP-100F series fermentation silicone defoaming agent) 1.6 L, mixing speed is 360 rpm, fermentation temperature is 36 ℃, fermentation time is 72 h, pH 6.8～7.2, the composition of described fermentation culture is: potato starch 8.5%, cottonseed protein 2.6%, K 2HPO 40.05%, NaCl 0.2%, MgSO 40.05%, CaCO 30.1%, pH7.2-7.4, potato starch, cottonseed protein, K 2HPO 4, NaCl, MgSO 4And CaCO 3The unit ratio of content is W/V;
    (D) metabolite extracting method in the NF0919 bacterial strain fermentation liquor
    Step (C) gained zymotic fluid is got supernatant at centrifugal 5 min of 10000 rpm, regulate pH to 2.0 with concentrated hydrochloric acid, precipitation occurs, again centrifugal, the collecting precipitation thing is used ethyl acetate extraction 3 times, and 35 ℃ of vacuum dryings get extract.
CN2012103546314A 2012-09-23 2012-09-23 Application of metabolins of streptomyces corchorusii NF0919 bacterial strain in prevention and control cucumber diseases Pending CN102870821A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103275892A (en) * 2013-05-14 2013-09-04 南京农业大学 Antagonism actinomycetes for preventing and killing continuous cropping cucumber fusarium wilt and microorganism organic fertilizer with antagonism actinomycetes
CN113174351A (en) * 2021-06-11 2021-07-27 成都云图控股股份有限公司 Disease-resistant growth-promoting mixed microbial flora

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4269971A (en) * 1978-12-29 1981-05-26 Taisho Pharmaceutical Co., Ltd. Antibiotic TM-531
CN101283684A (en) * 2007-11-19 2008-10-15 中国计量学院 Application of a compound for preventing and controlling cucumber fusarium wilt and tomato gray mold
CN101940211A (en) * 2010-03-19 2011-01-12 中国计量学院 Application of toyocamycin in preventing and curing cucumber wilt
CN102010835A (en) * 2010-07-27 2011-04-13 江苏丘陵地区镇江农业科学研究所 Streptomyces corchorusii strain NF0919, purpose and preparation method of active zymotic fluid thereof

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US4269971A (en) * 1978-12-29 1981-05-26 Taisho Pharmaceutical Co., Ltd. Antibiotic TM-531
CN101283684A (en) * 2007-11-19 2008-10-15 中国计量学院 Application of a compound for preventing and controlling cucumber fusarium wilt and tomato gray mold
CN101940211A (en) * 2010-03-19 2011-01-12 中国计量学院 Application of toyocamycin in preventing and curing cucumber wilt
CN102010835A (en) * 2010-07-27 2011-04-13 江苏丘陵地区镇江农业科学研究所 Streptomyces corchorusii strain NF0919, purpose and preparation method of active zymotic fluid thereof

Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN103275892A (en) * 2013-05-14 2013-09-04 南京农业大学 Antagonism actinomycetes for preventing and killing continuous cropping cucumber fusarium wilt and microorganism organic fertilizer with antagonism actinomycetes
CN113174351A (en) * 2021-06-11 2021-07-27 成都云图控股股份有限公司 Disease-resistant growth-promoting mixed microbial flora
CN113174351B (en) * 2021-06-11 2023-04-25 成都云图控股股份有限公司 Disease-resistant growth-promoting mixed microbial flora

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Application publication date: 20130116