CN105112295A - Fresh water fungus Arthrobotrys nonseptata and application thereof - Google Patents
Fresh water fungus Arthrobotrys nonseptata and application thereof Download PDFInfo
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- CN105112295A CN105112295A CN201510160375.9A CN201510160375A CN105112295A CN 105112295 A CN105112295 A CN 105112295A CN 201510160375 A CN201510160375 A CN 201510160375A CN 105112295 A CN105112295 A CN 105112295A
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Abstract
The invention relates to a fresh water fungus strain Arthrobotrys nonseptata YMF1.01852 and application thereof, belonging to the technical field of microbial pesticides. The fresh water fungus strain Arthrobotrys nonseptata YMF1.01852 is preserved in China General Microbiological Culture Collection Center, Institute of Microbiology, Chinese Academy of Sciences, No. 3, Yard 1, West Beichen Road, Chaoyang District, Beijing City, China, on December 10, 2014, with an accession number of CGMCC No. 10115. The fresh water fungus strain Arthrobotrys nonseptata YMF1.01852 is applied to preparation of biological agents used for preventing and treating root-knot nematode and Caenorhabditis elegans. The invention has the following advantages: the biological agents prepared from the fresh water fungus strain Arthrobotrys nonseptata YMF1.01852 have the characteristic of obvious nematicidal effect on root-knot nematode and Caenorhabditis elegans and have good application and development prospects.
Description
Technical field:
The invention belongs to microbial pesticide technical field, be specifically related to a strain fresh water fungi without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 bacterial strain and application thereof.
Background technology:
Plant nematode is one of important disease of plant, and the whole world is every year because the farm crop financial loss caused by plant nematode disease reaches 78,000,000,000 dollars.In China, nearly all crop such as plant nematode harm tobacco, flowers, vegetables, cotton, soybean, peanut, Chinese medicinal materials etc., becomes one of limiting factor important in agriculture production.About there is kind more than 3000 to the nematode of plant pest at present, mainly contain root knot nematode, Cyst nematode, pine wood nematode etc. in China.Root knot nematode agriculturally endangers a maximum class nematode, and according to estimates, in the total losses that whole world agriculture production causes because of all kinds of disaster every year, the harm of root knot nematode accounts for about 5% (1).
At present mainly chemical insecticide is relied on to the control of nematode, but life-time service chemical pesticide is except causing environmental pollution, remaining, destroying except the drawbacks such as the eubiosis, exacerbates the resistance of insect simultaneously.Utilize nemic natural enemy to have free from environmental pollution to carry out biological control, lasting medicine, insect such as not easily to develop immunity to drugs at the advantage, obtains each
government of stateattention.Finding from natural resource and find active nematicide metabolite, is the important channel (2) of development environment friendly GR.The aquatic fungi aboundresources of China, there is species diversity widely, and the research little (3 to its secondary metabolite, 4), the present invention is just based on above-mentioned theory, with root knot nematode and Caenorhabditis elegans for target, from fresh water fungus metabolite, finding efficient nematocidal active material, is research and development
novelnew resource found by nematocides.
Reference:
1.McCarterJP(2008)Nematology:terraincognitanomore.NatureBiotechnology26:882-884
2. the progress .20 (2) of Dong Jin gorgeous grade (2001) fungi nematicide metabolite
3.LuoJ,ZhangKQ,CaiL,KevinDH(2004)FreshwaterfungiinmainlandChina.23(1)
4. Dong Jin is gorgeous, the progress of Shen Kaize, Sun Rong (2011) fresh water fungal active metabolite.30(2)：206-217
Summary of the invention:
Order of the present invention is to provide a strain fresh water fungi without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 bacterial strain and application thereof.
The strain fresh water fungi that the present invention screens without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 bacterial strain on December 8th, 2014
preservationat Chinese microorganism strain
preservationmanagement
the councilcommon micro-organisms center;
preservationunit address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica;
preservationnumber: CGMCCNo.10115.
After fungi of the present invention is cultivated by test tube kind, then extract preparation after conventional liquid fermentation and culture, concrete steps are as follows:
1. test tube kind is cultivated
Culture medium prescription is potato agar (PDA) substratum, i.e. potato 200g; Glucose 20g; Agar 20g; Water 1000mL.
2. liquid fermentation and culture
Liquid fermentation medium is potato dextrose broth (PDB) substratum, i.e. potato 200g; Glucose 20g; Water 1000mL.
Test tube kind be inoculated in 500mL triangular flask (every bottled 150-250mL) liquid nutrient medium, at 25 ~ 28 DEG C, shaking speed is with 130 ~ 170rmin
-1, ferment and obtain fermented liquid in 10-16 days.
True fresh water fungi of the present invention prevents and treats the application in root knot nematode, Caenorhabditis elegans biotechnological formulation without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 bacterial strain in preparation.
The invention has the advantages that: feature root knot nematode and Caenorhabditis elegans without the biotechnological formulation prepared every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 bacterial strain with fresh water fungi to nematicide successful, has good application and development prospect.
Embodiment:
First to fresh water fungi without every Arthrobotrys (Arthrobotrysnonseptata)) YMF1.01852 bacterial strain carries out fermentation culture, then carries out eelworm-killing activity test to the meta-bolites after fermentation culture, determines its effect to nematode.
Embodiment 1: bacterial strain is without the cultivation every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852
To be inoculated on test tube nutrient agar without the mycelium every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852, culture medium prescription is PDA substratum, i.e. potato 200g; Glucose 20g; Agar 20g; Water 1000mL.Cultivate 8 days for 28 DEG C, obtain test tube kind.
Test tube kind be inoculated in 500mL triangular flask (every bottled 150mL) liquid nutrient medium, culture medium prescription is: potato 200g again; Glucose 20g; Water 1000mL, pH nature.Then, at 25 DEG C or 28 DEG C, shaking speed is with 130rmin
-1, or 170rmin
-1ferment and obtain fermented liquid in 10 days or 16 days.
Embodiment 2: bacterial strain is tested without the eelworm-killing activity every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852
Fermented liquid is concentrated on the basis of 5 times of liquid, successively dilution be 5 ×, 10 ×, 20 ×, 50 ×, respectively eelworm-killing activity test is carried out to root knot nematode, Caenorhabditis elegans.
1. prepare test preparation
Fermented liquid is concentrated on the basis of 5 times of liquid, successively dilution be 5 ×, 10 ×, 20 ×, 50 ×, respectively eelworm-killing activity test is carried out to root knot nematode, Caenorhabditis elegans.
2. prepare test and use nematode
A. root knot nematode is prepared
Tomato root is rinsed well, hatches in the sterilized water of 28 DEG C after picking pieces of an egg under the microscope.Adopt improvement modified Baermann funnel method to collect 2 instar larvaes: in funnel, to spread 60 object screen clothes, screen cloth repaves one deck medicated napkin, funnel bottom rubber hose is connected, and connect a blue electron gun head under rubber hose, rubber hose clip is clamped.After dull and stereotyped 2-3 time of aseptic water washing nematode, the nematode solution obtained is transferred in the screen cloth of 50um and is filtered, separation obtains the great-hearted nematode solution of tool (medicated napkin makes dead wire worm and impurity not to filter), unclamps clip and collects nematode, abandon supernatant after centrifugal after 4h.
B. Caenorhabditis elegans is prepared
Picking Caenorhabditis elegans culture, be seeded in the aseptic oat medium in triangular flask, cultivate 10d for 28 DEG C, when seeing that in substratum, a large amount of nematode is movable, picking culture of nematodes thing is put into 4 layers of lens wiping paper and is wrapped, put into the graceful funnel of the shellfish filling deionized water, according to motility and the action of gravity of himself of nematode, the test tube connected below funnel is utilized to collect nematode, this device leaves standstill 8-12h, collect the nematode be deposited on bottom test tube, nematode is sub-packed in the Ep pipe of 1.5mL, 5, 000rpm, supernatant is abandoned after centrifugal 30sec, and wash 3-5 time with sterilized water, concentrated for subsequent use.
3. without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 metabolite to the test of nematode effect
In 16 orifice plates through sterilizing, add that to be diluted successively by the fermented liquid of concentrated 5 times be 5 respectively ×, 10 ×, 20 ×, 50 × each 1mL of experiment fermented liquid, to add 1mL sterilized water in contrast, then 50 microlitre nematode suspension (containing nematode 96-100 bar) are respectively added respectively in process and contrast, put into 28 DEG C of thermostat containers to cultivate, observed once every 24 hours and record the mortality ratio of nematode, calculation correction mortality ratio is nematicide drug effect.Each process repetition 3 times.
Nemic death rate (%)=(verge of death borer population/confession examination nematode number) × 100
Corrected mortality (%)=[(process nemic death rate-contrast nemic death rate)/(1-contrasts nemic death rate)] × 100
A. without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 metabolite to the nematicide effect of Caenorhabditis elegans
Observe after 24 hours, find that most of nematode activity becomes slow.
After 48 hours observe, find most nematode dead, experimental data as
following table instituteshow:
Without every the different extension rate of Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 fermented liquid to the nematicide effect of Caenorhabditis elegans
Result shows, without having good eelworm-killing activity every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 meta-bolites to Caenorhabditis elegans.
B. without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 metabolite to the nematicide effect of root knot nematode
Observe after 24h, find that most nematode is dead, experimental data as
following table instituteshow:
Without every the different extension rate of Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 fermented liquid to the nematicide effect of Meloidogyne incognita
Result shows, without having good eelworm-killing activity every Arthrobotrys (Arthrobotrysnonseptata) meta-bolites to root knot nematode.
By showing the nematicide test-results of two kinds of nematodes, without being the fungi that a strain has using value every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852, demonstrate the application and development prospect that it is good.
Claims (2)
1. a strain fresh water fungi is without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 bacterial strain, preserving number: CGMCCNo.10115.
2. fresh water fungi according to claim 1 prevents and treats the application in root knot nematode, Caenorhabditis elegans biotechnological formulation without every Arthrobotrys (Arthrobotrysnonseptata) YMF1.01852 bacterial strain in preparation.
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| Application Number | Priority Date | Filing Date | Title |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108220207A (en) * | 2018-03-14 | 2018-06-29 | 陆平丰 | Compound microbial preparation and application thereof in preventing and treating root-knot nematodes |
| WO2020091031A1 (en) | 2018-11-02 | 2020-05-07 | 日本農薬株式会社 | Pest control agent composition and method for using same |
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|---|---|---|---|---|
| KR100791983B1 (en) * | 2007-04-17 | 2008-01-04 | (주) 카프코 | Microorganism of arthrobotrys sp. and microbial agent for preventing plant-parasitic nematodes comprising the same |
| WO2011099020A1 (en) * | 2010-02-09 | 2011-08-18 | Patel, Babubhai C. | Composition and method of preparation of fungal based product for controlling nematodes living in soil and damage to crops |
| CN101724569B (en) * | 2009-12-09 | 2011-12-21 | 云南大学 | Method for inducing nematode-trapping fungi to synchronously produce capturing organs |
| CN104293677A (en) * | 2014-04-08 | 2015-01-21 | 云南大学 | Arthrobotrys oligospora genetic engineering strain with nematode-killing function, and application thereof |
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2015
- 2015-04-07 CN CN201510160375.9A patent/CN105112295A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100791983B1 (en) * | 2007-04-17 | 2008-01-04 | (주) 카프코 | Microorganism of arthrobotrys sp. and microbial agent for preventing plant-parasitic nematodes comprising the same |
| CN101724569B (en) * | 2009-12-09 | 2011-12-21 | 云南大学 | Method for inducing nematode-trapping fungi to synchronously produce capturing organs |
| WO2011099020A1 (en) * | 2010-02-09 | 2011-08-18 | Patel, Babubhai C. | Composition and method of preparation of fungal based product for controlling nematodes living in soil and damage to crops |
| CN104293677A (en) * | 2014-04-08 | 2015-01-21 | 云南大学 | Arthrobotrys oligospora genetic engineering strain with nematode-killing function, and application thereof |
Non-Patent Citations (5)
| Title |
|---|
| HONGYAN SU 等: "Arthrobotrys latispora, a new nematode-trapping fungus from southwest China", 《MYCOTAXON》 * |
| SHIFU LI 等: "Arthrobotrys nonseptata, a new anamorph from an Orbilia species", 《MYCOTAXON》 * |
| ZHANG YING 等: "Genetic diversity and recombination in natural populations of the nematode-trapping fungus Arthrobotrys oligospora from China", 《ECOLOGY AND EVOLUTION》 * |
| 张克勤 等: "中国的捕食线虫真菌：V.节丛孢属", 《真菌学报》 * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108220207A (en) * | 2018-03-14 | 2018-06-29 | 陆平丰 | Compound microbial preparation and application thereof in preventing and treating root-knot nematodes |
| CN108220207B (en) * | 2018-03-14 | 2021-05-28 | 山东蒙德尔生物科技有限公司 | Compound microbial preparation and application thereof in preventing and treating root-knot nematodes |
| WO2020091031A1 (en) | 2018-11-02 | 2020-05-07 | 日本農薬株式会社 | Pest control agent composition and method for using same |
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Application publication date: 20151202 |