CN107841467A - A kind of biological pesticide for preventing and treating pine wood nematode and preparation method thereof - Google Patents
A kind of biological pesticide for preventing and treating pine wood nematode and preparation method thereof Download PDFInfo
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- CN107841467A CN107841467A CN201711141695.5A CN201711141695A CN107841467A CN 107841467 A CN107841467 A CN 107841467A CN 201711141695 A CN201711141695 A CN 201711141695A CN 107841467 A CN107841467 A CN 107841467A
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Abstract
The present invention relates to a kind of Paecilomyces lilacinus（Paecilomyces lilacinus）The anti-microbial inoculum of E22 bacterial strain fermentation liquor n-butanol extracts, its preparation method and the application in terms of pine wood nematode life is prevented and treated.Inclined-plane Paecilomyces lilacinus E22 culture parent species are prepared using Kolle flask, seed liquor is prepared, liquid fermentation is carried out using 500 mL triangular flasks, after zymotic fluid is filtered with sterile gauze after fermentation, centrifugation, by supernatant and n-butanol according to 1:1、1:2 and 1:3 ratio（Volume ratio）Mixing, is filtered, abandon water layer after static placement, and by n-butanol layer liquid, all evaporation is dry, and yellowish-brown powder wall built-up is arranged at round-bottomed flask bottom, obtains bacteriostatic active ingredients.Zymotic fluid n-butanol extract Tween 80 solution is dissolved, the 1/10 of original fermentation liquor filtered fluid volume is reverted to, is prepared into bacteria agent, carrying out indoor suppression nematode using the bacteria agent of preparation studies.
Description
Technical field
The present invention relates to a kind of biological pesticide for preventing and treating pine wood nematode, it is true that the agricultural chemicals is extracted from suppression pine wood nematode biological and ecological methods to prevent plant disease, pests, and erosion
Bacterium-Paecilomyces lilacinus（Paecilomyces lilacinus）Antibacterial metabolite in E22 zymotic fluids, further relates to and its prepares
Method, more particularly, to Paecilomyces lilacinus（Paecilomyces lilacinus）E22 liquid fermentation process flow and activity into
The separating and extracting process divided, and its application in terms of pine wood nematode is prevented and treated.
Background technology
Pine nematode（Bursaphelenchus xylophilus）It is the destructive disease of pine tree, it is dead after trees illness
It is high to die rate, difficulty of prevention and cure is big, can cause quite serious economic loss and very great ecocatastrophe, in economy of forestry and
There is very big threat in natural landscape, the title for having No.1 plant quarantine object in countries in the world.Paecilomyces lilacinus
（Paecilomyces lilacinus）Belong to endoparasitism fungi, be the Important Natural Enemy of plant nematode.For preventing and treating pine
Material nematode, always based on chemical agent, and the negative effect that chemical agent is brought so that the research of biological control turns into heat
Point.Fungal farm chemicals have the features such as toxicity is low, and selectivity is strong, highly-effective low-residue, and harm is low to caused by environment, both at home and abroad research
It is the Important Natural Enemy of Plant nematode to show Paecilomyces lilacinus.
Paecilomyces lilacinus（Paecilomyces lilacinus）In Ainsworth（1973）Belong to Fungi Imperfecti Asia in system
Door（Deuteromycotina）, Hyphomycetes（Hyphomycetes）, Hyphomycetales（Hyphoales）, hyphomycete section
（Hyphoaceae）, paecilomyces（Paecilomyces）.Paecilomyces lilacinus belongs to endoparasitism fungi, is plant nematode
Important Natural Enemy, easily culture, effect be high.It is therefore desirable to develop it is a kind of it is efficient, less toxic, non-harmful, stably, be easy to preserve
The anti-microbial inoculum of Paecilomyces lilacinus.
The content of the invention
In view of this, it is an object of the invention to provide a kind of biological pesticide and preparation method thereof for preventing and treating pine wood nematode,
And its application in terms of pine wood nematode is prevented and treated.
An aspect of of the present present invention, there is provided one plant have bacteriostatic activity Paecilomyces lilacinus (Paecilomyceslilacinus),
The bacterial strain is Paecilomyces lilacinus E22, is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC),
Preservation date is on 09 14th, 2017, and deposit number is CGMCC No：14621, depositary institution address is BeiJing, China.
A kind of another aspect of the present invention, there is provided biological pesticide for preventing and treating pine wood nematode, it is characterised in that the formulation of the agricultural chemicals
For pulvis, the active component of the pulvis is Paecilomyces lilacinus（Paecilomyces lilacinus）E22 zymotic fluid n-butanols carry
Take thing, the pulvis be by by filtering fermentation liquor, centrifugation, take supernatant, add n-butanol hybrid extraction, abandon water layer, by n-butanol
Layer is concentrated by evaporation what is obtained.
On the other hand, specific preparation technology is as follows：
（1）It is prepared by inclined-plane parent species：The Paecilomyces lilacinus that will have been activated（Paecilomyces lilacinus）E22 is inoculated into use gram
On improvement PDA inclined-planes prepared by family name's bottle, improvement PDA culture medium forms in proportion is：The g of potato 180 ~ 200, glucose 18 ~ 20
g、KH2PO4 2.5~3.5 g、MgSO4 1 ~ 1.5 g, the mL of water 800 ~ 1200, the constant temperature quiescent culture 4 ~ 6 under the conditions of 23 ~ 26 DEG C
After it, 1 × 10 is diluted to conidium under aseptic water washing5Individual spore/mL, 4 DEG C save backup.
（2）Paecilomyces lilacinus（Paecilomyces lilacinus）E22 liquid fermentation process flows：
Zymotic fluid culture medium prescription：The g/L of peptone 9 ~ 12, glucose 5 ~ 7 g/L, KH2PO4 9~11 g/L、CaCl2 0.7~
1.0g/L、MgSO4·7H2O 0.5~0.9 g/L、(NH4)2SO4 10~15 g/L、FeSO4·7H2O 4.0~6.0 mg/L、
ZnSO4·7H2O 1.2~1.6 mg/L、CoCl2·6H2O 3.5~4.0 mg/L、MnSO4·H2O 1.0~2.0 mg/L。
Fermentation process is fermented using 500 mL triangular flasks, and design parameter is as follows：
500 mL triangular flask zymotechniques：The mL of fermentation medium 200 ~ 250, the mL spore liquids of inoculum concentration 30 ~ 50, revolution 140 ~
160 rpm, initial pH 7.0, the d of cultivation cycle 5 ~ 7, solids content 5.0% ~ 6.0%.
（3）Paecilomyces lilacinus（Paecilomyces lilacinus）The extraction of E22 zymotic fluid bacteriostatic active ingredients and system
Standby technique
By Paecilomyces lilacinus（Paecilomyces lilacinus）After E22 zymotic fluids are filtered with sterile gauze, by supernatant with cold
Freeze the rpm of centrifuge 10000 ~ 12000 and centrifuge 10 ~ 15 min, by supernatant and n-butanol according to 1:1、1:2、1:3 ratio（Body
Product ratio）Mixing, at normal temperatures the h of 24 h of static placements ~ 48, separatory funnel filter, abandon water layer, by n-butanol layer with 45 ~ 50 DEG C
Rotary evaporation in vacuo concentrates, until all evaporation is dry for liquid, yellowish-brown powder wall built-up is arranged at round-bottomed flask bottom；Obtain biological pesticide
Pulvis.By Paecilomyces lilacinus（Paecilomyces lilacinus）Zymotic fluid n-butanol extract is with 0.5% ~ 2.0%（V/V）Tell
Warm 80 solution dissolving, reverts to the 1/10 of original fermentation liquor filtered fluid volume, is prepared into bacteria agent.
（4）Indoor suppression pine wood nematode research
Cultivate Botrytis cinerea：PDA culture medium is made first（Formula is same as above）.The ash for keeping laboratory in superclean bench
Grape spore connects bacterium to PDA culture medium, is cultivated a few days in 25 DEG C of constant incubators, waits Botrytis cinerea to cover with culture dish.
It is inoculated with pine wood nematode：The pine wood nematode that inoculation experiments room is kept in superclean bench is put in Botrytis cinerea
It is placed in a few days in 25 DEG C of constant incubators, waits nematode to eat up Botrytis cinerea.
Collect nematode liquid：Prepare one big funnel to be placed on iron stand, the rubber tube of one section of 10 cm length connect below funnel,
A tongs is filled on rubber tube, takes 4 layers of gauze to be placed on above big funnel, sterilized water is poured into and covers funnel edge.Will culture
Ware pine wood nematode solid-state PDA is all put into big funnel, puts 5-6 pieces, stands 24 h.After 24 h, with 30 mL centrifuge tubes from only
The sterilized water in big funnel is connect at water folder, centrifuge is used after having connect, in centrifuge（3500 r/min, 5 min）In
After impurity is removed in centrifugation, nematode liquid is diluted with PBST buffer solutions, repeats 2 to 3 times, is finally collected into nematode liquid.
Kill pine wood nematode toxicity test in interior：The μ L of active component 150 are taken using liquid-transfering gun, the μ L of nematode liquid 50, are mixed
Amount to being positioned in 96 orifice plates, be repeated 3 times per sample, extract 50 μ L with liquid-transfering gun in each orifice plate every time, and show using optics
Micro mirror is measured and records pine wood nematode death condition, and time interval is 24 h.Utilize the h numbers of 24 h of record, 48 h, 72
According to analyzing and draw a conclusion.
It is an advantage of the invention that：The present invention is the preparation side of antibacterial metabolite microbial inoculum in Paecilomyces lilacinus E22 zymotic fluids
Method, the raw material used is abundance, inexpensive for reproducible fungal resources, has significant inhibition to pine wood nematode, this
Invention has the characteristics that efficient, less toxic, nuisanceless, and pine wilt disease caused by pine wood nematode is with a wide range of applications.
Embodiments of the invention are described in further detail below embodiment.
The Paecilomyces lilacinus of embodiment 1（Paecilomyces lilacinus）The system of active ingredients Biogenic microbial inoculum in E22 zymotic fluids
Standby technique：
（1）It is prepared by inclined-plane parent species：Will be in PDA plate culture medium（The mm of diameter 90）The upper Paecilomyces lilacinus activated
（Paecilomyces lilacinus）E22 is inoculated on the improvement PDA inclined-planes prepared with Kolle flask（Improvement PDA culture medium is pressed
Ratio forms：The g of potato 200, glucose 20 g, KH2PO4 3.0 g、MgSO4 1.5 g, water 1000 mL, 121 DEG C, 0.15
25 min of Mpa sterilizings）, the constant temperature quiescent culture after 5 days under the conditions of 25 DEG C, 1 is diluted to conidium under aseptic water washing ×
105Individual spore/ml, 4 DEG C save backup.
（2）Paecilomyces lilacinus（Paecilomyces lilacinus）E22 liquid fermentation process flows：
Zymotic fluid culture medium prescription：
The g/L of peptone 9, glucose 5 g/L, KH2PO4 9 g/L、CaCl2 0.7 g/L、MgSO4·7H2O 0.5 g/L、
(NH4)2SO4 10 g/L、FeSO4·7H2O 4.0 mg/L、ZnSO4·7H2O 1.2 mg/L、CoCl2·6H2O 3.5 mg/L、
MnSO4·H2O 1.0 mg/L。
500 mL triangular flask zymotechniques：The mL of fermentation medium 200, the mL spore liquids of inoculum concentration 30, the rpm of revolution 140,
Initial pH 7.0, the d of cultivation cycle 5, solids content 5.0% ~ 6.0%.
（3）Paecilomyces lilacinus（Paecilomyces lilacinus）The extraction of E22 zymotic fluid bacteriostatic active ingredients and system
Standby technique
By Paecilomyces lilacinus（Paecilomyces lilacinus）After E22 zymotic fluids are filtered with sterile gauze, by supernatant with cold
Freeze the rpm of centrifuge 10000 and centrifuge 10 min, by supernatant and n-butanol according to 1:3 ratio（Volume ratio）Mixing, in normal temperature
Under 48 h of static placement, separatory funnel filtering, abandon water layer, n-butanol layer concentrate with 45 DEG C of rotary evaporation in vacuo, up to liquid
All evaporation is dry, and yellowish-brown powder wall built-up is arranged at round-bottomed flask bottom；Obtain bacteriostatic active ingredients.By Paecilomyces lilacinus
（Paecilomyces lilacinus）E22 zymotic fluids n-butanol extract is dissolved with 0.5% Tween 80 solution, is reverted to primary
The 1/10 of zymotic fluid filtered fluid volume, is prepared into bacteria agent.
（4）Indoor suppression pine wood nematode research
Kill pine wood nematode toxicity test in interior：The μ L of active component 140, the μ L of nematode liquid 50 are taken using liquid-transfering gun, is mixed altogether
With being positioned in 96 orifice plates, it is repeated 3 times per sample, extracts 50 μ L with liquid-transfering gun in each orifice plate every time, and use light microscope
It is measured and records pine wood nematode death condition, time interval is 24 h.Utilize the h of 24 h of record, 48 h, 72 data point
Analyse and draw a conclusion.
Indoor measurement result:The E22 insecticidal effects of extracting n-butyl alcohol are fairly obvious, extracting n-butyl alcohol ratio 1:1,1:2,
1:3 have all reached 100% in the h corrected mortalities of 24 h, 48 h, 72, and n-butanol extracts as extractant to Paecilomyces lilacinus E22
Take effect very notable, the insecticide active substance in E22 can all be extracted substantially, so as to reach efficient desinsection effect
Fruit.
The Paecilomyces lilacinus of embodiment 2（Paecilomyces lilacinus）Active ingredients Biogenic microbial inoculum in E22 zymotic fluids
Preparation technology：
（1）It is prepared by inclined-plane parent species：Will be in PDA plate culture medium（The mm of diameter 90）The upper Paecilomyces lilacinus activated
（Paecilomyces lilacinus）E22 is inoculated on the improvement PDA inclined-planes prepared with Kolle flask（Improvement PDA culture medium is pressed
Ratio forms：The g of potato 200, glucose 20 g, KH2PO4 3.0 g、MgSO4 1.5 g, water 1000 mL, 121 DEG C, 0.15
25 min of Mpa sterilizings）, the constant temperature quiescent culture after 5 days under the conditions of 25 DEG C, 1 is diluted to conidium under aseptic water washing ×
105Individual spore/ml, 4 DEG C save backup.
（2）Paecilomyces lilacinus（Paecilomyces lilacinus）E22 liquid fermentation process flows：
Zymotic fluid culture medium prescription：
The g/L of peptone 9, glucose 5 g/L, KH2PO4 9 g/L、CaCl2 0.7 g/L、MgSO4·7H2O 0.5 g/L、
(NH4)2SO4 10 g/L、FeSO4·7H2O 4.0 mg/L、ZnSO4·7H2O 1.2 mg/L、CoCl2·6H2O 3.5 mg/L、
MnSO4·H2O 1.0 mg/L。
500 mL triangular flask zymotechniques：The mL of fermentation medium 250, the mL spore liquids of inoculum concentration 50, the rpm of revolution 160,
Initial pH 7.0, the d of cultivation cycle 7, solids content 5.0% ~ 6.0%.
（3）Paecilomyces lilacinus（Paecilomyces lilacinus）The extraction of E22 zymotic fluid bacteriostatic active ingredients and system
Standby technique
By Paecilomyces lilacinus（Paecilomyces lilacinus）After E22 zymotic fluids are filtered with sterile gauze, by supernatant with cold
Freeze the rpm of centrifuge 10000 and centrifuge 15 min, by supernatant and n-butanol according to 1:2 ratio（Volume ratio）Mixing, in normal temperature
Under 48 h of static placement, separatory funnel filtering, abandon water layer, n-butanol layer concentrate with 45 DEG C of rotary evaporation in vacuo, up to liquid
All evaporation is dry, and yellowish-brown powder wall built-up is arranged at round-bottomed flask bottom；Obtain bacteriostatic active ingredients.By Paecilomyces lilacinus
（Paecilomyces lilacinus）E22 zymotic fluids n-butanol extract is dissolved with 0.5% Tween 80 solution, is reverted to primary
The 1/10 of zymotic fluid filtered fluid volume, is prepared into bacteria agent.
（4）Indoor suppression pine wood nematode research
Kill pine wood nematode toxicity test in interior：The μ L of active component 150, the μ L of nematode liquid 50 are taken using liquid-transfering gun, is mixed altogether
With being positioned in 96 orifice plates, it is repeated 3 times per sample, extracts 50 μ L with liquid-transfering gun in each orifice plate every time, and use light microscope
It is measured and records pine wood nematode death condition, time interval is 24 h.Utilize the h of 24 h of record, 48 h, 72 data point
Analyse and draw a conclusion.
Indoor measurement result:The E22 insecticidal effects of extracting n-butyl alcohol are fairly obvious, extracting n-butyl alcohol ratio 1:1,1:2,
1:3 have all reached 100% in the h corrected mortalities of 24 h, 48 h, 72, and n-butanol extracts as extractant to Paecilomyces lilacinus E22
Take effect very notable, the insecticide active substance in E22 can all be extracted substantially, nematode effect is killed so as to reach efficient
Fruit.
Claims (12)
1. one plant of Paecilomyces lilacinus (Paecilomyces lilacinus) with bacteriostatic activity, it is characterised in that the bacterial strain is
Paecilomyces lilacinus E22, it is deposited in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC), preservation day
Phase is on 09 14th, 2017, and deposit number is CGMCC No：14621.
A kind of 2. biological pesticide for preventing and treating pine wood nematode, it is characterised in that the formulation of the agricultural chemicals be pulvis, the pulvis it is active into
It is divided into Paecilomyces lilacinus E22 zymotic fluid n-butanol extracts.
3. biological pesticide as claimed in claim 2, it is characterised in that the pulvis is by by Paecilomyces lilacinus E22 zymotic fluids
Filtering, centrifugation, take supernatant, add n-butanol hybrid extraction, abandon water layer, n-butanol layer is concentrated by evaporation into what is obtained.
4. biological pesticide as claimed in claim 2, it is characterised in that the pulvis can be used for preventing after the dissolving of Tween 80 solution
Control pine wood nematode.
5. biological pesticide as claimed in claim 4, it is characterised in that the concentration of the Tween 80 is 0.5%~2.0% (V/
V)。
6. biological pesticide as claimed in claim 4, it is characterised in that obtained volume of the pulvis after Tween 80 dissolves
For the 1/10 of original fermentation liquor filtered fluid volume.
7. application of the Paecilomyces lilacinus E22 zymotic fluids n-butanol extract in terms of pine wood nematode is prevented and treated.
8. a kind of preparation method for the biological pesticide for preventing and treating pine wood nematode, it is characterised in that this method is by Paecilomyces lilacinus E22
Filtering fermentation liquor, centrifugation, take supernatant, add n-butanol hybrid extraction, abandon water layer, n-butanol layer are concentrated by evaporation to obtain biological agriculture
Medicine powder.
9. preparation method as claimed in claim 8, it is characterised in that the inclined-plane parent species of the Paecilomyces lilacinus E22 by with
It is prepared by lower step：The Paecilomyces lilacinus E22 activated is inoculated on the improvement PDA inclined-planes prepared with Kolle flask, improvement PDA trainings
Foster base forms in proportion is：180~200g of potato, 18~20g of glucose, KH2PO42.5~3.5g, MgSO41~1.5g,
800~1200mL of water, constant temperature quiescent culture is diluted after 4~6 days with conidium under aseptic water washing under the conditions of 23~26 DEG C
To 1 × 105Individual spore/mL, 4 DEG C save backup.
10. preparation method as claimed in claim 8, it is characterised in that the technological process of the Paecilomyces lilacinus E22 zymotic fluids
It is as follows：
Zymotic fluid culture medium prescription：9~12g/L of peptone, 5~7g/L of glucose, 9~11g/L of KH2PO4, CaCl2 0.7~
0.5~0.9g/L of 1.0g/L, MgSO47H2O, 10~15g/L of (NH4) 2SO4,4.0~6.0mg/L of FeSO47H2O,
1.2~1.6mg/L of ZnSO47H2O, 3.5~4.0mg/L of CoCl26H2O, 1.0~2.0mg/L of MnSO4H2O,
Fermentation process is fermented using 500mL triangular flasks, and design parameter is as follows：
500mL triangular flask zymotechniques：200~250mL of fermentation medium, inoculum concentration 30~50mL spore liquids, revolution 140~
160rpm, initial pH 7.0,5~7d of cultivation cycle, solids content 5.0%~6.0%.
11. preparation method as claimed in claim 8, it is characterised in that the Paecilomyces lilacinus E22 zymotic fluid sterile gauzes
After filtering, supernatant is centrifuged into 10~15min with 10000~12000rpm of refrigerated centrifuge, by supernatant and n-butanol according to 1:
1、1:2、1:3 ratio (volume ratio) mixes, at normal temperatures static placement 24h~48h, and separatory funnel filtering, abandon water layer will just
Butanol layer is concentrated with 45~50 DEG C of rotary evaporation in vacuo, until all evaporation is dry for liquid, yellowish-brown powder is arranged at round-bottomed flask bottom
Wall built-up；Obtain biological pesticide pulvis.
12. preparation method as claimed in claim 11, it is characterised in that by the biological pesticide pulvis with 0.5%~2.0%
(V/V) Tween 80 solution dissolves, and the 1/10 of original fermentation liquor filtered fluid volume is reverted to, for preventing and treating pine wood nematode.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102657235A (en) * | 2012-05-10 | 2012-09-12 | 肇庆市真格生物科技有限公司 | Nematicidal composition containing verticillium chlamydosporium and Paecilomyces lilacinus and preparation method thereof |
| CN102657155A (en) * | 2012-05-10 | 2012-09-12 | 肇庆市真格生物科技有限公司 | Nematicide granules with sawdust as carrier and preparation method thereof |
| CN106342882A (en) * | 2016-08-25 | 2017-01-25 | 山东省联合农药工业有限公司 | Nematicidal composition and application thereof |
-
2017
- 2017-11-16 CN CN201711141695.5A patent/CN107841467A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102657235A (en) * | 2012-05-10 | 2012-09-12 | 肇庆市真格生物科技有限公司 | Nematicidal composition containing verticillium chlamydosporium and Paecilomyces lilacinus and preparation method thereof |
| CN102657155A (en) * | 2012-05-10 | 2012-09-12 | 肇庆市真格生物科技有限公司 | Nematicide granules with sawdust as carrier and preparation method thereof |
| CN106342882A (en) * | 2016-08-25 | 2017-01-25 | 山东省联合农药工业有限公司 | Nematicidal composition and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| 李国红等: "一种新的杀线虫担子菌", 《云南大学学报 (自然科学版) 》 * |
| 李萍等: "放线菌 BJLSH9 菌株兼抗线虫及烟草疫霉菌的生防活性及其杀线虫代谢产物鉴定", 《云南大学学报 (自然科学版)》 * |
| 魏博涵等: "15 个生防菌株乙酸乙酯提取物对松材线虫的杀虫活性", 《林业科技》 * |
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