CN106987539A - A kind of zymotic fluids of bacterium Lysobacter soli 6262 and its application - Google Patents
A kind of zymotic fluids of bacterium Lysobacter soli 6262 and its application Download PDFInfo
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- CN106987539A CN106987539A CN201710265851.2A CN201710265851A CN106987539A CN 106987539 A CN106987539 A CN 106987539A CN 201710265851 A CN201710265851 A CN 201710265851A CN 106987539 A CN106987539 A CN 106987539A
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- 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
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Abstract
A kind of zymotic fluids of bacterium Lysobacter soli 6262 and its application.The zymotic fluids of bacterium Lysobacter soli 6262 are screened through bacterium bacterial strain and the preparation process of the zymotic fluids of Lysobacter soli 6262 is obtained.Bacterium bacterial strain Lysobacter soli 6262 are deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number on January 09th, 2017：CGMCC No.13555.The zymotic fluids of bacterium Lysobacter soli 6262 are applied in poisoning Meloidogyne incognita (Meloidogyne incognita) and Caenorhabditis elegans (Caenorhabditis elegans) biological agent is prepared.Beneficial effect is：It is high-efficiency low-toxicity, free from environmental pollution, the biological pesticide preparation for preparing poisoning Meloidogyne incognita (M.incognita) and Caenorhabditis elegans (C.elegans) can be used for.
Description
Technical field
The present invention relates to a kind of zymotic fluids of bacterium Lysobacter soli 6262 and its application, microorganism belonging to genus biological pesticide
Technical field.
Background technology
Disease caused by Plant nematode turns into the second major class disease in agricultural production, the warp caused every year to the whole world
Ji loss is up to 157,000,000,000 dollars (Abad et al., 2008).It is prevented and treated based on chemical pesticide, with long-term use variousization
The problem of environmental pollution that medicine of learning to farm is brought is on the rise, and part chemical nematicides are disabled, therefore utilize nemic natural enemy
Biological control is carried out to nematode to be increasingly taken seriously.Microorganism is the main groups in nemic natural enemy, its active secondary metabolism
Product is the basis for finding biological pesticide and new drug.Wherein bacterium due to species it is various, as find eelworm-killing activity secondary generation
Thank to one of important sources of product.
The bacterium bacterial strain that the present invention is used is Lysobacter soli 6262, and there is the zymotic fluid of the bacterium poisoning nematode to live
Property.Retrieved through domestic and foreign literature, have no and disclose report with identical of the present invention.
The content of the invention
It is an object of the invention to provide a kind of zymotic fluids of bacterium Lysobacter soli 6262 and its application.
What the present invention was realized in：
1. the bacterium bacterial strain Lysobacter soli 6262 that the present invention is used are deposited in China on January 09th, 2017
Microbiological Culture Collection administration committee common micro-organisms center；Depositary institution address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City
No. 3 Institute of Microorganism, Academia Sinica；Preserving number：CGMCC No.13555.
2. the preparation of the zymotic fluids of bacterium Lysobacter soli 6262
Culture medium：NB culture mediums are conventional medium.The composition and compound method of culture medium are as follows:
NB culture mediums：Peptone 10.0g, beef extract 3.0g and NaCl 5.0g are weighed, with deionized water dissolving and is settled to
1L, adjusts pH to 7.2, and 121 DEG C sterilize 20 minutes.
100 μ L Lysobacter soli 6262,25 are inoculated with the triangular flask equipped with 100mL NB fluid nutrient mediums
Culture -6 days 3 days in DEG C of -38 DEG C constant-temperature tables.Zymotic fluid supercentrifuge centrifuged under 10000rpm 5min separation bacterium solution and
Thalline, Aspirate supernatant is placed on standby in -4 DEG C of refrigerator, obtains the active-fermented broth that uses of the present invention.
3. Caenorhabditis elegans (Caenorhabditis elegans) and Meloidogyne incognita (Meloidogyne
Incognita culture)
Oat medium is cultivated for Caenorhabditis elegans (C.elegans)；Oatmeal 10.0g, water 10mL infiltrate oat
Piece, 120 DEG C of high-temperature sterilization 20min.Inoculation nematode be positioned in 28 DEG C of incubator cultivate 7 days or 10 days after, be placed in 4 DEG C of refrigerators
In save backup.Nematode suspension is made of sterilized water when using standby (1000 every milliliter or so).
Tomato Root System is used for the preparation of Meloidogyne incognita (M.incognita).Morbidity Tomato Root System is taken, is put into water and soaks
Bubble 3 times, every time at least 10 hours.Soak centrifuges 2min through 2000rpm, and abandoning supernatant adds 494g/L sucrose, after mixing
2000rpm centrifuges 2min, and supernatant is cleaned with the bolter of 800 mesh, collects the larva on bolter and worm's ovum.Washing lotion is used again
100th, the bolter of 200 and 500 mesh is rinsed repeatedly, and it is worm's ovum to collect gleanings-major part on 500 mesh bolters；500 mesh point
From the filtered solution on sieve, the bolter cleaning through 800 mesh again, collects the instar larvae of polypide -2 on bolter.Separated worm and worm
Ovum is stored in standby in 4 DEG C of refrigerator.
4. experimental method
Liquid infusion method：The suspension of 4.5mL test samples solution, 500 μ L about containing 500 nematodes is taken in a diameter of 6cm
Culture dish in, gently mix after culture dish room temperature is placed.Randomly choosed under anatomical lens the timing of 10 visuals field (1 hour, 2
Hour, 8 hours, 24 hours, 48 hours) observed, count disease pest number (being used as standard so that nematode is motionless) and bus borer population.Often
Individual test sample sets three repetitions, and using the NB culture mediums of non-inoculating strain as control, obtains test sample to the lethal of nematode
Rate, with virulence of the fatal rate height evaluation sample to nematode.
The zymotic fluids of bacterium Lysobacter soli 6262 of the present invention are preparing poisoning Meloidogyne incognita
(M.incognita) and in Caenorhabditis elegans (C.elegans) biological agent apply.
The beneficial effects of the present invention are：High-efficiency low-toxicity, free from environmental pollution, fermenation raw liquid is at 8 hours to beautiful hidden bar line
The fatal rate of worm (C.elegans) reaches 100%, reached in 48 hours fatal rates to Meloidogyne incognita (M.incognita)
To more than 90%.It can be used in preparing poisoning Meloidogyne incognita (M.incognita) and Caenorhabditis elegans (C.elegans)
Biological agent.
Embodiment：
The present invention is described in further detail with embodiment below, but present disclosure is not limited thereto.In embodiment
The preparation processes of the zymotic fluids of bacterium Lysobacter soli 6262 and poisoning Meloidogyne incognita (M.incognita) and beautiful
Description of the experimental method of hidden rhabditida (C.elegans) with Summary.
Embodiment one：
The μ L of bacterium Lysobacter soli 6,262 100 after activation are inoculated in (250mL tri- in liquid NB culture mediums
The bottled 100mL culture mediums in angle), in 25 DEG C, cultivated 3 days under the conditions of 180rpm.After nutrient solution 10000rpm is centrifuged 5 minutes, take
Supernatant is used as test sample.It the results are shown in Table 1.
Poisoning result (%) of the zymotic fluids of 1 bacterium Lysobacter soli of table 6262 to C.elegans nematodes
As a result show：The zymotic fluids of bacterium Lysobacter soli 6262 have pole to Caenorhabditis elegans (C.elegans)
Strong toxic action, fermenation raw liquid reached 100% in 8 hours fatal rates to Caenorhabditis elegans (C.elegans).
Embodiment two：
The present embodiment is basically identical using identical experiment medicament, experimental method with embodiment one, only will be for examination nematode
Replace with Meloidogyne incognita.
The μ L of bacterium Lysobacter soli 6,262 100 after activation are inoculated in (250mL tri- in liquid NB culture mediums
The bottled 100mL culture mediums in angle), in 30 DEG C, cultivated 5 days under the conditions of 180rpm.After nutrient solution 10000rpm is centrifuged 5 minutes, take
Supernatant is used as test sample.It the results are shown in Table 2.
Poisoning knot of the zymotic fluids of 2 bacterium Lysobacter soli of table 6262 to Meloidogyne incognita (M.incognita)
Really (%)
As a result show：Bacterium Lysobacter soli 6262 cultivate the zymotic fluid pair of 5 days in 30 DEG C, under the conditions of 180rpm
Meloidogyne incognita (M.incognita) has strong toxic action, and fermenation raw liquid is at 48 hours to Meloidogyne incognita
(M.incognita) fatal rate reaches more than 90%.
Embodiment three：
The present embodiment is basically identical using identical experiment medicament, experimental method with embodiment one, only will be for examination nematode
Replace with Meloidogyne incognita.
The μ L of bacterium Lysobacter soli 6,262 100 after activation are inoculated in (250mL tri- in liquid NB culture mediums
The bottled 100mL culture mediums in angle), in 38 DEG C, cultivated 6 days under the conditions of 180rpm.After nutrient solution 10000rpm is centrifuged 5 minutes, take
Supernatant is used as test sample.It the results are shown in Table 3.
Poisoning knot of the zymotic fluids of 3 bacterium Lysobacter soli of table 6262 to Meloidogyne incognita (M.incognita)
Really (%)
As a result show：Bacterium Lysobacter soli 6262 cultivate the zymotic fluid pair of 6 days in 38 DEG C, under the conditions of 180rpm
Meloidogyne incognita (M.incognita) has strong toxic action, and fermenation raw liquid is at 48 hours to Meloidogyne incognita
(M.incognita) fatal rate reaches more than 90%.
Claims (2)
1. a kind of zymotic fluids of bacterium Lysobacter soli 6262, it is characterised in that bacterium Lysobacter soli 6262 are sent out
Zymotic fluid is obtained by the following steps：
A. bacterium Lysobacter soli 6262 are deposited in Chinese microorganism strain preservation management committee on January 09th, 2017
Member can common micro-organisms center；Depositary institution address：Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Chinese Academy of Sciences microorganism
Research institute；Preserving number：CGMCC No.13555；
B. the preparation of the zymotic fluids of bacterium Lysobacter soli 6262
NB culture mediums：Peptone 10.0g, beef extract 3.0g and NaCl 5.0g are weighed, with deionized water dissolving and 1L is settled to,
PH to 7.2 is adjusted, 121 DEG C sterilize 20 minutes；
It is inoculated with 100 μ L bacterium Lysobacter soli6262 in the triangular flask equipped with 100mL NB fluid nutrient mediums, 25 DEG C-
Culture -6 days 3 days in 38 DEG C of constant-temperature tables；Zymotic fluid supercentrifuge centrifuges 5min separation bacterium solutions and bacterium under 10000rpm
Body, Aspirate supernatant is placed on standby in -4 DEG C of refrigerator, obtains the active-fermented broth that uses of the present invention.
2. the zymotic fluids of bacterium Lysobacter soli 6262 described in claim 1 are preparing poisoning Meloidogyne incognita
(M.incognita) and in Caenorhabditis elegans (C.elegans) biological agent apply.
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| CN201710265851.2A CN106987539B (en) | 2017-04-21 | 2017-04-21 | A kind of 6262 fermentation liquid of bacterium Lysobacter soli and its application |
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| CN201710265851.2A CN106987539B (en) | 2017-04-21 | 2017-04-21 | A kind of 6262 fermentation liquid of bacterium Lysobacter soli and its application |
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| CN106987539B CN106987539B (en) | 2019-08-02 |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110283756A (en) * | 2019-07-18 | 2019-09-27 | 云南大学 | A kind of preparation method and application of the molten bacillus fermentation liquid of glue |
| CN113817630A (en) * | 2021-08-04 | 2021-12-21 | 云南大学 | Roseburia sp strain and application thereof |
| CN114480164A (en) * | 2021-12-22 | 2022-05-13 | 沣田宝农业科技有限公司 | Lysobacter tumefaciens and application thereof |
Citations (2)
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|---|---|---|---|---|
| CN101177671A (en) * | 2007-12-05 | 2008-05-14 | 南京农业大学 | Biocontrol bacteria strain preventing and curing plant disease |
| CN105145633A (en) * | 2015-07-31 | 2015-12-16 | 北京泰克美高新技术有限公司 | Drug controlling plant nematodes |
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2017
- 2017-04-21 CN CN201710265851.2A patent/CN106987539B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101177671A (en) * | 2007-12-05 | 2008-05-14 | 南京农业大学 | Biocontrol bacteria strain preventing and curing plant disease |
| CN105145633A (en) * | 2015-07-31 | 2015-12-16 | 北京泰克美高新技术有限公司 | Drug controlling plant nematodes |
Non-Patent Citations (1)
| Title |
|---|
| MYUNG KYUM KIM等: "Solirubrobacter soli sp. nov., isolated from soil of a ginseng field", 《INTERNATIONAL JOURNAL OF SYSTEMATIC AND EVOLUTIONARY MICROBIOLOGY》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110283756A (en) * | 2019-07-18 | 2019-09-27 | 云南大学 | A kind of preparation method and application of the molten bacillus fermentation liquid of glue |
| CN110283756B (en) * | 2019-07-18 | 2022-11-11 | 云南大学 | Preparation method and application of colloidal lysobacter fermentation broth |
| CN113817630A (en) * | 2021-08-04 | 2021-12-21 | 云南大学 | Roseburia sp strain and application thereof |
| CN113817630B (en) * | 2021-08-04 | 2022-02-25 | 云南大学 | Roseburia sp strain and application thereof |
| CN114480164A (en) * | 2021-12-22 | 2022-05-13 | 沣田宝农业科技有限公司 | Lysobacter tumefaciens and application thereof |
| CN114480164B (en) * | 2021-12-22 | 2023-08-22 | 沣田宝农业科技有限公司 | Lysobacter agrobacterium and application thereof |
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