CN101100646A - Paecilomyces lilacinus and application thereof - Google Patents
Paecilomyces lilacinus and application thereof Download PDFInfo
- Publication number
- CN101100646A CN101100646A CNA2007101178181A CN200710117818A CN101100646A CN 101100646 A CN101100646 A CN 101100646A CN A2007101178181 A CNA2007101178181 A CN A2007101178181A CN 200710117818 A CN200710117818 A CN 200710117818A CN 101100646 A CN101100646 A CN 101100646A
- Authority
- CN
- China
- Prior art keywords
- yes
- lilacinus
- bacterium
- paecilomyces lilacinus
- cgmcc
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
A (P.lilacinus)YES-2 CGMCC No. 2012, its bacterial agent and use are disclosed. The process is carried out by taking polymer sodium alginate as base skeleton, taking (P.lilacinus)YES-2 CGMCC No. 2012 mycelium and spore mixture as active ingredients, taking diatomite as carrier, forming sodium alginate and polyvalent positive ion into gel and sodium alginate-calcium chloride reacting to obtain final product. The prevention rate reaches to 50-60%. It's stable, cheap and convenient. It has no chemical residual and pollution for agricultural product and environment. It can be used to prevent plant root nematode.
Description
Technical field
The present invention relates to a Paecilomyces lilacinus and application, particularly relate to the application in control plant root-knot nematode disease of a Paecilomyces lilacinus and microbial inoculum thereof and this microbial inoculum.
Background technology
In the four big cause of diseases that cause plant infectious diseases, the harm of plant nematode surpasses bacterium and virus, is only second to fungal disease.Plant nematode is caused about 1,000 hundred million dollars of the loss that world agriculture produces every year, wherein root knot nematode (Meloidogyne spp.) is the maximum class plant nematode of harm, and the annual financial loss that only important in the world cash crop is caused is just up to tens billion of dollars.In Meloidogyne (Meloidogyne), root knot nematode disease generation with Meloidogyne incognita (M.incognita), peanut root-knot nematode (M.arenaria), javanese root knot nematode (M.javanica) and four kinds of northern root knot nematode (M.hapla) is the most general, and is also maximum to the harm of farm crop.The reproductivity of root knot nematode is extremely strong, and each female worm can lay eggs more than 1000 pieces usually.Root knot nematode does not have tangible host's specialization usually, the plant that can infect is above 3000 kinds, adhere to 114 sections separately, comprise monocotyledons, dicotyledons, herbaceous plant and xylophyta all can be caused heavy losses to most food crop, oil crops, fibre crops, tobacco, tealeaves, fruit tree, vegetables, medicinal material and flowers etc.After being subjected to its harm, crop quality descends, the underproduction significantly, even total crop failure, and the generation of isochrone parasitosis has aggravated the harm of soil-borne diseases such as blight, verticillium and damping-off again, causes the plant resistivity to descend, and infected by other pathogen.
In the method for such disease of control, because root knot nematode survives in the soil usually and roots of plants in, so general chemical pesticide is difficult to control its harm, so such disease becomes the frequent object that uses of a large amount of highly toxic pesticide, and the security of agricultural-food production is constituted a serious threat.Simultaneously, because chemical nematode killing agent can only be controlled nematode population in a short time, also there is the cost height simultaneously, holds problems such as imitating short and resistance.In addition, the method for preventing and treating that some are traditional, though can play certain preventive and therapeutic effect as crop rotation, plantation disease-resistant variety etc., crop that can crop rotation in actual production is also few, the crop varieties that root knot nematode is had resistance is very limited.Therefore, traditional control method can not solve such disease.In sum, because lack practical, efficient, continuable improvement technology at present, the root knot nematode disease generation area constantly enlarges, the damage to crops kind continues to increase, cause agriculture production to face a severe challenge, the wireworm-killing biologic agricultural chemicals has boundless using value and market outlook.
Safe and effective control root knot nematode class disease is the pressing problem that needs to be resolved hurrily in the present agriculture production, and measures such as existing chemical prevention can not be satisfied this requirement, and therefore utilizing nematode biological and ecological methods to prevent plant disease, pests, and erosion resource that it is regulated and control is the inexorable trend of Future Development.Current, along with the raising of people's environmental protection consciousness with to the continuous increase of " non-harmful product " demand, biocontrol fungi demonstrates more and more important effect in the plant nematode diseases biological control.But must be pointed out, only have seldom a part of bacterial strain to be used to research and develop biological prevention and control agent, and these microbial inoculums really play preventive and therapeutic effect seldom on producing although occurring in nature has numerous fungus and nematode closely related.In addition, still there is certain shortcoming in the nematode biological prevention and control agent at present, has limited its application greatly, is mainly reflected in:
1. the biological prevention and control agent of existing control plant nematode diseases mostly is spore preparation, and is higher to upstream activeconstituents production requirement, often needs two-step fermentation (liquid fermenting is in conjunction with solid fermentation) could obtain activeconstituents one fungal spore, and preparation technology is very loaded down with trivial details.
2. the formulation level is low, control of nematode at present mostly is pulvis, suspension agent etc. with biological prevention and control agent, technology is extensive, soil is poor for applicability, be unfavorable for that edatope uses, and often be difficult to overcome the adverse environmental factors of antibacterial sprouting in the soil, be difficult for forming colony, and the many forms with kind of clothing agent or seed dressing of these preparations occur, and it is very limited to take content of molds.Thereby the shortage market competitiveness, its application is restricted.
3. poor stability, the forfeiture of microbial inoculum vigor is very rapid, causes the biological prevention and control agent shelf-lives short, is difficult to commercially produce.In order to guarantee that preparation has the shelf-lives of long period, often strict preservation condition has directly limited the large-scale application of Mycophyta biological pesticide.
In sum, although plant nematode diseases biological control type agricultural chemicals has the boundless market space in the soil, obtained some by biological control research for a long time and had the bacterial strain of certain biological and ecological methods to prevent plant disease, pests, and erosion potentiality plant nematode, but its commercial application is but made slow progress, and the preparation that can satisfy nematode biological and ecological methods to prevent plant disease, pests, and erosion needs in actual production lacks very much.Therefore existing biological and ecological methods to prevent plant disease, pests, and erosion resource will promote the biological control work of plant nematode disease to jump onto a new step in case have breakthrough in that formulation is technical greatly.
Summary of the invention
The purpose of this invention is to provide a strain has preventive and therapeutic effect to the plant root-knot nematode disease Paecilomyces lilacinus bacterium.
Provided by the present invention is Paecilomyces lilacinus bacterium (Paecilomyces.lilacinus) YES-2, this bacterial strain is preserved in Da Tun road, Chaoyang District, BeiJing, China China Committee for Culture Collection of Microorganisms common micro-organisms center on April 20th, 2007, and deposit number is CGMCC NO.2012.
Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 bacterial strain has following feature: bacterium colony felted, surface just are white, become the grape wine redness when producing spore, and sporulation quantity are many more, and color is dark more, and the back side is white in color or burgundy.This bacterial strain is gone up growth comparatively fast at malt extract medium (MEA), also can grow on the PDA substratum.Begin to produce spore after cultivating for 1 week, its conidiophore is upright, 2-4 of verticillate arrangement bottle stalk on the falx, the top conidium of concatenating.Conidium is ellipsoid or fusiform, and outer wall is smooth, and size is (2.0-3.0) * (1.5-2.2) μ m.The temperature range of this strain growth is 8-38 ℃, and the best is 25-30 ℃.This bacterial strain all can be grown between pH2-11, and the appropriate pH value is 5-9.
Second purpose of the present invention provides the cultural method of a kind of above-mentioned Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCCNo.2012.
The cultural method of Paecilomyces lilacinus bacterium provided by the present invention (P.lilacinus) YES-2 CGMCC No.2012, be that Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 is inoculated in Paecilomyces lilacinus bacterium (P.lilacinus) the YES-2 CGMCC No.2012 dedicated liquid substratum, cultivate down at 23-25 ℃, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 obtains enlarged culturing; The prescription of described Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 dedicated liquid substratum is: maltose (C source) 20-40g (being preferably 30g), yeast (N source) 5-7g (being preferably 6.12g), K
2HPO
4(0.5-2g being preferably 1g), MgSO
4(0.1-1g being preferably 0.5g), KCl 0.1-1g (being preferably 0.5g), FeSO
4(0.001-0.05g being preferably 0.01g), ZnSO
47H
2O5-20mg (being preferably 10mg), water is settled to 1L, pH6-7.
For improving the growth velocity of thalline, can carry out shaking culture to this bacterium, vibration velocity is preferably 120-180rpm.
Described incubation time is generally 120-180h.
For obtaining higher growth velocity, the cultural method of described Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 also is included in the preceding step of bacterial classification being carried out the test tube activation culture of inoculation, method can be: Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 is seeded on the potato agar substratum, cultivate down at 25-27 ℃, obtain activated Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012; Described potato agar culture medium prescription is: potato 200g, and glucose 20g, agar 16g, water is settled to 1L, about pH6-7.
The test tube activation culture time of described Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 can be 72-96h.
The 3rd purpose of the present invention provides a kind of Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 microbial inoculum.
Paecilomyces lilacinus bacterium provided by the present invention (P.lilacinus) YES-2 CGMCC No.2012 microbial inoculum, its activeconstituents are Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012.
For obtaining better stability, described microbial inoculum can adopt the microballoon formulation, and its composition of raw materials can comprise following component (every 100g):
Sodium alginate 1-4g,
Diatomite 8-15g,
Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 1-10g,
Water adds water to 100g.
For the performance that makes microbial inoculum is more complete, can also comprise the calcium chloride of 1-3g in the above-mentioned microbial inoculum
More preferably the composition of raw materials of Paecilomyces lilacinus bacterium (P.lilacinus) the YES-2 CGMCC No.2012 microbial inoculum of microballoon formulation can be two kinds of situations, and first kind is:
Sodium alginate 1-4g,
Diatomite 10-14g,
Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 5-10g,
Water 72-84g,
Calcium chloride 1-2g.
Wherein, the weight addition of sodium alginate, diatomite, Paecilomyces lilacinus bacterium YES-2 and water should equal 100g.
Second kind of situation is:
Sodium alginate 1-3g,
Diatomite 12-15g,
Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 1-5g,
Water 77-86g,
Calcium chloride 1-2.5g.
Wherein, the weight addition of sodium alginate, diatomite, Paecilomyces lilacinus bacterium YES-2 and water should equal 100g.
Water in the composition of raw materials of above-mentioned microballoon formulation Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 microbial inoculum is preferably sterilized water or distilled water.
Another object of the present invention provides a kind of method for preparing Paecilomyces lilacinus bacterium (P.lilacinus) the YES-2 CGMCC No.2012 microbial inoculum of above-mentioned microballoon formulation.
Preparation method provided by the present invention can may further comprise the steps:
1) gets material by following composition of raw materials: sodium alginate 1-4g, diatomite 8-15g, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 1-10g, water 71-90g, calcium chloride 1-3g; Wherein, the weight addition of sodium alginate, diatomite, Paecilomyces lilacinus bacterium YES～2 and water should equal 100g;
2) preparation of microballoon sodium alginate gel liquid: sodium alginate is mixed with sterilized water, heat also constantly stirring down at 40-60 ℃ it is dissolved, obtain sodium alginate aqueous solution, cooling, standby;
3) Paecilomyces lilacinus bacterium YES-2 and diatomite are joined step 2) preparation sodium alginate aqueous solution in, mixing;
4) preparation CaCl
2Solution: water preparation volumetric molar concentration is the calcium chloride solution of 0.1-0.3M, and is standby;
5) mixing solutions of sodium alginate that step 3) is obtained and Paecilomyces lilacinus bacterium YES-2 adds in the container, instils under constant hydraulics, and the sodium alginate micro ball latax that forms instiling falls into the CaCl of step 4) preparation
2In the solution, after sodium alginate micro ball and the calcium chloride solution reaction, generate calcium alginate microsphere;
6) the calcium alginate microsphere hardening that step 5) obtained stirs, then calcium alginate microsphere leached, and flushing, drying obtains the Paecilomyces lilacinus bacterium YES-2 microbial inoculum of microballoon formulation.
In the preparation method of above-mentioned Paecilomyces lilacinus bacterium YES-2 microbial inoculum, the composition of raw materials in the step 1) is preferably (every 100g): sodium alginate 1-4g, and diatomite 10-14g, Paecilomyces lilacinus bacterium YES-2 5-10g adds water to 100g, is instilled into calcium chloride 1-2g again; Perhaps be: sodium alginate 1-3g, diatomite 12-15g, Paecilomyces lilacinus bacterium YES-21-5g adds water to 100g, is instilled into calcium chloride 1-2.5g again.
Step 2) preferably sodium alginate aqueous solution is cooled to below 30 ℃ in.
Paecilomyces lilacinus bacterium YES-2 in the step 3) can be the mixture of its mycelium and spore thereof.
Hardening churning time in the step 6) can be 30-60 minute; Available sterilized water cleans calcium alginate microsphere; Described drying means can be vacuum-drying or dries in the shade.
Used water is preferably sterilized water or distilled water in the above-mentioned microballoon formulation Paecilomyces lilacinus bacterium YES-2 bacterial preparation process.
The invention provides a strain has higher insecticidal activity to plant nematode one root knot nematode root knot nematode fungi autoeciousness bacterial strain Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 and microbial inoculum thereof.This microbial inoculum is to be basic framework with the macromolecular material sodium alginate, mycelium, spore mixture with Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 are activeconstituents, with diatomite is carrier, characteristic according to sodium alginate and polyvalent cation formation gel, adopt the method for sodium alginate-calcium chloride reaction, the wireworm-killing biologic bacterial agent that applies usefulness is delayed in having of being prepared from.Experiment showed, that this microbial inoculum has nematicide effect preferably, control rate can reach 50-60%, and it is comparatively stable, do not have deleterious chemical residue (nontoxic, noresidue), do not pollute agricultural-food and environment (not with environment in other substance reaction), cost performance is high and use characteristics easily.The present invention will play a significant role in the control of plant root-knot nematode disease, have a extensive future.
Below in conjunction with specific embodiment the present invention is described in further details.
Description of drawings
Fig. 1 is for being the aspect graph of the microspheric biocontrol fungicide of activeconstituents with Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012
Embodiment
Method therefor is ordinary method if no special instructions among the following embodiment, and all percentage concentrations are mass percent concentration if no special instructions, and the solvent in all substratum is water.
Separation and the preservation of embodiment 1, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012
Gather E Shanta pasture, Yunnan tobacco root knot nematode old complaint, with tap water flushing old complaint 3-5 minute to remove soil, then old complaint is placed under the anatomical lens, choose egg capsule with pincet, handle egg capsule 1 minute with to egg capsule surface sterilization and release wire worm's ovum with 1% clorox, clorox ovum suspension crossed 200 μ m and 30 μ m mesh screens respectively, with sterile purified water flushing 3 times to remove clorox, approximately per 100 ovum are coated on the PDA flat board, observe bacterium colony every day and situation occurs.Finally on the tobacco root-knot nematode egg, separate and obtain a strain nematicidal fungi bacterial strain, called after YES-2, be preserved in the China Committee for Culture Collection of Microorganisms common micro-organisms center that is positioned at Da Tun road, Chaoyang District, BeiJing, China on April 20th, 2007, deposit number is CGMCC NO.2012.Through identifying that this bacterial strain is the Paecilomyces lilacinus bacterium, has following feature: bacterium colony felted, surface just are white, become the grape wine redness when producing spore, and sporulation quantity are many more, and color is dark more, and the back side is white in color or burgundy.This bacterial strain is gone up growth comparatively fast at malt extract medium (MEA), also can grow on the PDA substratum.Begin to produce spore after cultivating for 1 week, its conidiophore is upright, 2-4 of verticillate arrangement bottle stalk on the falx, the top conidium of concatenating.Conidium is ellipsoid or fusiform, and outer wall is smooth, and size is (2.0-3.0) * (1.5-2.2) μ m.The temperature range of this strain growth is 8-38 ℃, and the best is 25-30 ℃.This bacterial strain all can be grown between pH2-11, and the appropriate pH value is 5-9.
The cultivation of embodiment 2, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 and the preparation of microbial inoculum thereof
One, the cultivation of Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012
1, the test tube kind is cultivated
The potato agar culture medium prescription: potato 200g, glucose 20g, agar 16g, water is settled to 1L, pH6-7.
Paecilomyces lilacinus bacterium YES-2 is seeded on the potato agar substratum in the test tube, cultivates 72-96h down, obtain activated Paecilomyces lilacinus bacterium YES-2 at 25 ℃.
2, enlarged culturing
Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 dedicated liquid culture medium prescription: maltose (C source) 30g, yeast (N source) 6.12g, K
2HPO
41g, MgSO
40.5g, KCl0.5g, FeSO
40.01g, ZnSO
47H
2O 10mg, water is settled to 1L, pH6-7.
The Paecilomyces lilacinus bacterium YES-2 that step 1 is activated is inoculated in the Paecilomyces lilacinus bacterium YES-2 dedicated liquid substratum, cultivates 120h under 25 ℃, 150rpm, and Paecilomyces lilacinus bacterium YES-2 obtains enlarged culturing.
Two, the preparation of Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 microspheric microbial inoculum
Be equipped with microspheric Paecilomyces lilacinus bacterium of the present invention (P.lilacinus) YES-2 CGMCC No.2012 microbial inoculum in order to the below legal system, concrete preparation method may further comprise the steps:
1) takes by weighing raw material: sodium alginate 20kg, diatomite 100kg, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2CGMCC No.2012 (mixture of mycelium and spore thereof) 10kg, sterilized water 870kg, calcium chloride 22kg;
2) preparation of microballoon sodium alginate gel liquid: sodium alginate is mixed with sterilized water, heat also constantly stirring down at 40 ℃ it is dissolved, obtain sodium alginate aqueous solution, be cooled to about 30 ℃, standby;
3) Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 liquid culture filter is collected back homogenate and is smashed, centrifugally collect the back and diatomite joins step 2 jointly) in the sodium alginate aqueous solution of preparation, abundant stirring and evenly mixing, standby;
4) preparation CaCl
2Solution: be the calcium chloride solution of 0.2M with sterilized water preparation volumetric molar concentration, standby;
5) mixing solutions of sodium alginate that step 3) is obtained and Paecilomyces lilacinus bacterium YES-2 adds pressurization instillation in the airtight pressure vessel, and the sodium alginate micro ball latax that forms instiling falls into the CaCl of step 4) preparation
2In the solution, after sodium alginate micro ball and the calcium chloride solution reaction, generate calcium alginate microsphere;
6) the calcium alginate microsphere hardening that step 5) is obtained stirred after 60 minutes, leached microballoon, used aseptic water washing 3 times, and microballoon is dry under vacuum condition, obtained the Paecilomyces lilacinus bacterium YES-2 microbial inoculum of microballoon formulation.Form as shown in Figure 1, the heavy 0.022-0.024 gram of simple grain microballoon, diameter 4.8 ± 0.2mm, after testing, the CFU/g of Paecilomyces lilacinus bacterium YES-2 is 10
7-10
8
The cultivation of embodiment 3, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 and the preparation of microbial inoculum thereof
One, the cultivation of Paecilomyces lilacinus bacterium YES-2
1, the test tube kind is cultivated
The potato agar culture medium prescription: potato 200g, glucose 20g, agar 16g, water is settled to 1L, pH6-7.
Paecilomyces lilacinus bacterium YES-2 is inoculated on the potato agar substratum in the test tube, cultivates 72h down, obtain activated Paecilomyces lilacinus bacterium YES-2 at 25 ℃.
2, enlarged culturing
Paecilomyces lilacinus bacterium YES-2 dedicated liquid culture medium prescription: maltose 20g, yeast 5g, K
2HPO
40.5g, MgSO
40.1g, KCl0.1g, FeSO
40.001g, ZnSO
47H
2O5mg, water is settled to 1L, pH6-7.
The Paecilomyces lilacinus bacterium YES-2 that step 1 is activated is inoculated in the Paecilomyces lilacinus bacterium YES-2 dedicated liquid substratum, cultivates 150h under 26 ℃, 180rpm, and Paecilomyces lilacinus bacterium YES-2 obtains enlarged culturing.
Two, the preparation of Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 microspheric microbial inoculum
Be equipped with microspheric Paecilomyces lilacinus bacterium YES-2 microbial inoculum of the present invention in order to the below legal system, concrete preparation method may further comprise the steps:
1) takes by weighing raw material: sodium alginate 10kg, diatomite 150kg, Paecilomyces lilacinus bacterium YES-2 (mixture of mycelium and spore thereof) 20kg, calcium chloride 11kg, sterilized water 820kg;
2) preparation of microballoon sodium alginate gel liquid: sodium alginate is mixed with sterilized water, heat also constantly stirring down at 50 ℃ it is dissolved, obtain sodium alginate aqueous solution, be cooled to about 30 ℃, standby;
3) Paecilomyces lilacinus bacterium YES-2 homogenate is smashed, centrifugally collects the back and diatomite joins step 2 jointly) in the sodium alginate aqueous solution of preparation, abundant stirring and evenly mixing, standby;
4) preparation CaCl
2Solution: be the calcium chloride solution of 0.1M with sterilized water preparation volumetric molar concentration, standby;
5) mixing solutions of sodium alginate that step 3) is obtained and Paecilomyces lilacinus bacterium YES-2 adds in the airtight pressure vessel and instils under constant hydraulics, and the sodium alginate micro ball latax that forms instiling falls into the CaCl of step 4) preparation
2In the solution, after sodium alginate micro ball and the calcium chloride solution reaction, generate calcium alginate microsphere;
6) the calcium alginate microsphere hardening that step 5) is obtained stirred 30 minutes, then calcium alginate microsphere is leached, with aseptic water washing 2 times, leach microballoon vacuum-drying, obtain Paecilomyces lilacinus bacterium (P.lilacinus) the YES-2 CGMCC No.2012 microbial inoculum of microballoon formulation.The microbial inoculum form is identical with the microbial inoculum of embodiment 1 preparation, the heavy 0.032-0.034 gram of simple grain microballoon, and diameter 4.5 ± 0.3mm, after testing, the CFU/g of Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCCNo.2012 is 10
7-10
8
The cultivation of embodiment 4, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 and the preparation of microbial inoculum thereof
One, the cultivation of Paecilomyces lilacinus bacterium YES-2
1, the test tube kind is cultivated
The potato agar culture medium prescription: potato 200g, glucose 20g, agar 16g, water is settled to 1L, pH6-7.
Paecilomyces lilacinus bacterium YES-2 is seeded on the potato agar substratum in the test tube, cultivates 72h down, obtain activated Paecilomyces lilacinus bacterium YES-2 at 28 ℃.
2, enlarged culturing
Paecilomyces lilacinus bacterium YES-2 CGMCC dedicated liquid culture medium prescription: maltose 40g, yeast 7g, K
2HPO
42g, MgSO
41g, KCl1g, FeSO
40.05g, ZnSO
47H
2O 20mg, water is settled to 1L, pH6-7.
The Paecilomyces lilacinus bacterium YES-2 that step 1 is activated is inoculated in the Paecilomyces lilacinus bacterium YES-2 dedicated liquid substratum, cultivates 180h under 23 ℃, 120rpm, and Paecilomyces lilacinus bacterium YES-2 obtains enlarged culturing.
Two, the preparation of Paecilomyces lilacinus bacterium YES-2 microspheric microbial inoculum
Be equipped with microspheric Paecilomyces lilacinus bacterium YES-2 microbial inoculum of the present invention in order to the below legal system, concrete preparation method may further comprise the steps:
1) takes by weighing raw material: sodium alginate 20kg, diatomite 80kg, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2CGMCC No.2012 (mixture of mycelium and spore thereof) 30kg, calcium chloride 33kg, sterilized water 870kg;
2) preparation of microballoon sodium alginate gel liquid: sodium alginate is mixed with sterilized water, heat also constantly stirring down at 60 ℃ it is dissolved, obtain sodium alginate aqueous solution, be cooled to about 30 ℃, standby;
3) Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 homogenate is smashed, centrifugally collects the back and diatomite joins step 2 jointly) in the sodium alginate aqueous solution of preparation, abundant stirring and evenly mixing, standby;
4) preparation CaCl
2Solution: be the calcium chloride solution of 0.3M with sterilized water preparation volumetric molar concentration, standby;
5) mixing solutions of sodium alginate that step 3) is obtained and Paecilomyces lilacinus bacterium YES-2 adds in the airtight pressure vessel and instils under constant hydraulics, and the sodium alginate micro ball latax that forms instiling falls into the CaCl of step 4) preparation
2In the solution, after sodium alginate micro ball and the calcium chloride solution reaction, generate calcium alginate microsphere;
6) the calcium alginate microsphere hardening that step 5) is obtained stirred 45 minutes, then calcium alginate microsphere was leached, and used aseptic water washing 4 times, dried in the shade after leaching, and obtained Paecilomyces lilacinus bacterium (P.lilacinus) the YES-2 CGMCCNo.2012 microbial inoculum of microballoon formulation.The microbial inoculum form is identical with the microbial inoculum of embodiment 1 preparation, the heavy 0.018-0.020 gram of simple grain microballoon, and diameter 4.7 ± 0.2mm, after testing, the CFU/g of Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 is 10
7-10
8
The cultivation of embodiment 5, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 and the preparation of microbial inoculum thereof
One, the cultivation of Paecilomyces lilacinus bacterium YES-2
1, the test tube kind is cultivated
The potato agar culture medium prescription: potato 200g, glucose 20g, agar 16g, water is settled to 1L, pH6-7.
Paecilomyces lilacinus bacterium YES-2 is seeded on the potato agar substratum in the test tube, cultivates 96h down, obtain activated Paecilomyces lilacinus bacterium YES-2 at 25 ℃.
2, enlarged culturing
Paecilomyces lilacinus bacterium YES-2 dedicated liquid culture medium prescription: maltose 25g, yeast 6.5g, K
2HPO
41.5g, MgSO
40.8g, KCl0.4g, FeSO
40.03g, ZnSO
47H
2O10mg, water is settled to 1L, pH6-7.
The Paecilomyces lilacinus bacterium YES-2 that step 1 is activated is inoculated in the Paecilomyces lilacinus bacterium YES-2 dedicated liquid substratum, cultivates 120h under 25 ℃, 150rpm, and Paecilomyces lilacinus bacterium YES-2 obtains enlarged culturing.
Two, the preparation of Paecilomyces lilacinus bacterium YES-2 microspheric microbial inoculum
Be equipped with microspheric Paecilomyces lilacinus bacterium YES-2 microbial inoculum of the present invention in order to the below legal system, concrete preparation method may further comprise the steps:
1) takes by weighing raw material: sodium alginate 25kg, diatomite 120kg, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2CGMCC No.2012 (mixture of mycelium and spore thereof) 25kg, calcium chloride 11kg, sterilized water 830kg;
2) preparation of microballoon sodium alginate gel liquid: sodium alginate is mixed with sterilized water, heat also constantly stirring down at 45 ℃ it is dissolved, obtain sodium alginate aqueous solution, be cooled to about 30 ℃, standby;
3) Paecilomyces lilacinus bacterium YES-2 homogenate is smashed, centrifugally collects the back and diatomite joins step 2 jointly) in the sodium alginate aqueous solution of preparation, abundant stirring and evenly mixing, standby;
4) preparation CaCl
2Solution: be the calcium chloride solution of 0.1M with sterilized water preparation volumetric molar concentration, standby;
5) mixing solutions of sodium alginate that step 3) is obtained and Paecilomyces lilacinus bacterium YES-2 adds in the airtight pressure vessel and instils under constant hydraulics, and the sodium alginate micro ball latax that forms instiling falls into the CaCl of step 4) preparation
2In the solution, after sodium alginate micro ball and the calcium chloride solution reaction, generate calcium alginate microsphere;
6) the calcium alginate microsphere hardening that step 5) is obtained stirred 50 minutes, then calcium alginate microsphere was leached, and used aseptic water washing 4 times, dried in the shade after the filtration, obtained the Paecilomyces lilacinus bacterium YES-2 microbial inoculum of microballoon formulation.The microbial inoculum form is identical with the microbial inoculum of embodiment 1 preparation, the heavy 0.026-0.028 gram of simple grain microballoon, and diameter 4.6 ± 0.2mm, after testing, the CFU/g of Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 is 10
7-10
8
The biocontrol effect of embodiment 6, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 microbial inoculum detects
Paecilomyces lilacinus bacterium (P.lilacinus) the YES-2CGMCC No.2012 microbial inoculum of microballoon formulation that detects embodiment 2 preparations with following experiment is to the prevention effect of root knot nematode (is example with the cucumber root knot nematode), and experimental technique is as follows:
Selecting green a kind of jade plantation center, the serious Mentougou, Beijing City of root knot nematode disease morbidity vegetable greenhouse booth is booth, and the kind plant is a cucumber, and kind is " Diana " mini gherkin.Experiment sub-district area is 20 square metres, and every processing repeats for three times, and the sub-district is arranged as the random alignment method.When cucumber seedling is transplanted, reagent agent is adopted the method spread manuer in holes, impose under the root of the crop, experimental session is managed routinely, the time be the mid-September-late January next year.Reagent agent is as follows:
Handle 1: blank (not dispenser),
Handle 2: the biological nematicide microballoon of the present invention microbial inoculum 2kg/ mu,
Handle 3: the biological nematicide microballoon of the present invention microbial inoculum 4kg/ mu,
Handle 4: the biological nematicide microballoon of the present invention microbial inoculum 8kg/ mu,
Behind crop harvesting, dig out every processing Radix Cucumidis sativi, investigate according to root knot nematode classification investigation index, specific as follows: as 0 grade=do not have root knot, on the root of 1 grade of 1-10% root knot to be arranged, 2 grades of 11-20%, 3 grades of 21-30%, 4 grades of 31-40%, 5 grades of 41-50%, 6 grades of 51-60%, 7 grades of 61-70%, 8 grades of 71-80%, 9 grades of 81-90%, 10 grades of 91-100%.
The prevention effect calculation formula is: prevention effect=(contrast disease index-processing disease index)/contrast disease index.Wherein, the embodiment experimental result is as shown in table 1, as can be seen, using after Paecilomyces lilacinus bacterium of the present invention (P.lilacinus) the YES-2 CGMCC No.2012 microbial inoculum all has in various degree preventive effect to root knot nematode, compare with blank and can significantly reduce root knot nematode harm, and prevention effect improves constantly with the increase of amount of application.
Table 1 Paecilomyces lilacinus bacterium of the present invention YES-2 microbial inoculum is to the prevention effect detected result of cucumber root knot nematode
| Disease index | Prevention effect | |
| Handle 1 (contrast) | 74.13 | - |
| Embodiment 2 handles 2 embodiment 2 and handles 3 embodiment, 2 processing, 4 embodiment, 2 processing 5 | 62.17 54.33 34.67 19.33 | 16.13 26.71 53.23 73.92 |
The microbial inoculum that utilizes embodiment 3-5 to be obtained is according to the method described above tested, and the effect that obtains is consistent with the microbial inoculum that embodiment 2 is obtained.
Claims (10)
1, a Paecilomyces lilacinus (P.lilacinus) YES-2, its biological preserving number is CGMCC NO.2012.
2, the method for the described Paecilomyces lilacinus bacterium of a kind of cultivation claim 1 (P.lilacinus) YES-2 CGMCC No.2012, be that Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 is inoculated in Paecilomyces lilacinus bacterium (P.lilacinus) the YES-2 CGMCC No.2012 dedicated liquid substratum, cultivate down at 25-30 ℃, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 obtains enlarged culturing; The prescription of described Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 dedicated liquid substratum is: maltose 20-40g, yeast 5-7g, K
2HPO
40.5-2g, MgSO
40.1-1g, KCl 0.1-1g, FeSO
40.001-0.05g, ZnSO
47H
2O5-20mg, water is settled to 1L, pH6-7.
3, cultural method according to claim 2 is characterized in that: vibrate in the described culturing process, vibration velocity is 120-180rpm; Incubation time is 120-180h; Before the inoculation, earlier Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 bacterial classification is carried out the test tube activation culture, method is: Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 is seeded on the potato agar substratum, cultivate down at 25-30 ℃, obtain activated Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012; The prescription of described potato agar substratum is: potato 200g, and glucose 20g, agar 16g, water is settled to 1L, pH6-7.
4, a kind of Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 microbial inoculum, its activeconstituents are the described Paecilomyces lilacinus bacterium of claim 1 (P.lilacinus) YES-2 CGMCC No.2012.
5, microbial inoculum according to claim 4 is characterized in that: described microbial inoculum is the microballoon formulation, and its composition of raw materials comprises following component:
Sodium alginate 1-4g,
Diatomite 8-15g,
Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 1-10g,
Water adds water to 100g.
6, microbial inoculum according to claim 5 is characterized in that: the composition of raw materials that also comprises the calcium chloride of 1-3g in the described microbial inoculum comprises following component:
7, microbial inoculum according to claim 6 is characterized in that: the raw material of described microbial inoculum is composed of the following components:
Sodium alginate 1-4g,
Diatomite 10-14g,
Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 5-10g,
Water 72-84g,
Calcium chloride 1-2g.
Wherein, the weight addition of sodium alginate, diatomite, Paecilomyces lilacinus bacterium YES-2 and water should equal 100g.
8, microbial inoculum according to claim 6 is characterized in that: the raw material of described microbial inoculum is composed of the following components:
Sodium alginate 1-3g,
Diatomite 12-15g,
Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 1-5g,
Water 77-86g,
Calcium chloride 1-2.5g.
Wherein, the weight addition of sodium alginate, diatomite, Paecilomyces lilacinus bacterium YES-2 and water should equal 100g
9, a kind of method for preparing the described Paecilomyces lilacinus bacterium of claim 6 (P.lilacinus) YES-2 CGMCC No.2012 microbial inoculum may further comprise the steps:
1) gets material by following composition of raw materials: sodium alginate 1-4g, diatomite 8-15g, Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCC No.2012 1-10g, water 71-90g, calcium chloride 1-3g; Wherein, the weight addition of sodium alginate, diatomite, Paecilomyces lilacinus bacterium YES-2 and water should equal 100g;
2) preparation of microballoon sodium alginate gel liquid: sodium alginate is mixed with sterilized water, heat also constantly stirring down at 40-60 ℃ it is dissolved, obtain sodium alginate aqueous solution, cooling, standby;
3) Paecilomyces lilacinus bacterium YES-2 and diatomite are joined step 2) preparation sodium alginate aqueous solution in, mixing;
4) preparation CaCl
2Solution: water preparation volumetric molar concentration is the calcium chloride solution of 0.1-0.3M, and is standby;
5) mixing solutions of sodium alginate that step 3) is obtained and Paecilomyces lilacinus bacterium YES-2 adds in the container, instils under constant hydraulics, and the sodium alginate micro ball latax that forms instiling falls into the CaCl of step 4) preparation
2In the solution, after sodium alginate micro ball and the calcium chloride solution reaction, generate calcium alginate microsphere;
6) the calcium alginate microsphere hardening that step 5) obtained stirs, then calcium alginate microsphere leached, and flushing, drying obtains the Paecilomyces lilacinus bacterium YES-2 microbial inoculum of microballoon formulation.
10, the application of any described Paecilomyces lilacinus bacterium (P.lilacinus) YES-2 CGMCCNo.2012 microbial inoculum in control plant root-knot nematode disease among the claim 4-8.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101178181A CN101100646A (en) | 2007-06-25 | 2007-06-25 | Paecilomyces lilacinus and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007101178181A CN101100646A (en) | 2007-06-25 | 2007-06-25 | Paecilomyces lilacinus and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN101100646A true CN101100646A (en) | 2008-01-09 |
Family
ID=39035095
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNA2007101178181A Pending CN101100646A (en) | 2007-06-25 | 2007-06-25 | Paecilomyces lilacinus and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN101100646A (en) |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20110300606A1 (en) * | 2010-06-04 | 2011-12-08 | Chih-Hui Yang | Formula of medium for culturing cordyceps spp. |
| CN102286421A (en) * | 2011-09-16 | 2011-12-21 | 中国林业科学研究院森林生态环境与保护研究所 | Liquid fermentation culture method for paecilomyces lilacinus |
| CN102293220A (en) * | 2011-09-20 | 2011-12-28 | 中国农业科学院农业环境与可持续发展研究所 | Nanocrystallized synergistic rodenticide |
| CN102382775A (en) * | 2011-11-18 | 2012-03-21 | 中国科学院微生物研究所 | Eelworm killing metarhizium and application thereof |
| CN102742607A (en) * | 2012-07-23 | 2012-10-24 | 中国农业大学 | Paecilomyces lilacinus fungal spore oil suspending agent and preparation method as well as application thereof |
| CN102851219A (en) * | 2012-09-25 | 2013-01-02 | 中国热带农业科学院环境与植物保护研究所 | Paecilomyces lilacinus and application thereof |
| TWI385248B (en) * | 2010-05-05 | 2013-02-11 | Univ Ishou | A formula of culturing medium for cordyceps spp. |
| CN102925405A (en) * | 2012-05-29 | 2013-02-13 | 中国林业科学研究院森林生态环境与保护研究所 | Preparation method of paecilomyces lilacinus spore powder |
| CN103589649A (en) * | 2013-11-08 | 2014-02-19 | 湖北森源生态科技股份有限公司 | Production method of agricultural microbial paecilomyces lilacinus for nematode prevention and control |
| CN103749547A (en) * | 2013-12-25 | 2014-04-30 | 浙江农林大学 | Entomophthorales water floating type granules and application thereof |
| CN106376602A (en) * | 2016-08-31 | 2017-02-08 | 江西天人生态股份有限公司 | Paecilomyces lilacinus granule and preparation method thereof |
| CN106879607A (en) * | 2015-12-15 | 2017-06-23 | 内蒙古农业大学 | Prevent and treat the biological pesticide of apple tree canker |
| CN107586737A (en) * | 2017-09-18 | 2018-01-16 | 华中农业大学 | The Bei Laisi bacillus of one plant of anti-yellowing cucurbit wilt and its Microencapsulated Slow microbial inoculum |
| CN108294050A (en) * | 2018-03-16 | 2018-07-20 | 佛山市普尔玛农化有限公司 | Including Paecilomyces lilacinus kills line composition |
| CN110878257A (en) * | 2019-10-09 | 2020-03-13 | 北京四纬生物科技有限公司 | Paecilomyces lilacinus culture method and application |
| CN112111480A (en) * | 2020-10-14 | 2020-12-22 | 南宁汉和生物科技股份有限公司 | Method for prolonging shelf life of paecilomyces lilacinus by using double-layer embedding technology |
-
2007
- 2007-06-25 CN CNA2007101178181A patent/CN101100646A/en active Pending
Cited By (21)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| TWI385248B (en) * | 2010-05-05 | 2013-02-11 | Univ Ishou | A formula of culturing medium for cordyceps spp. |
| US20110300606A1 (en) * | 2010-06-04 | 2011-12-08 | Chih-Hui Yang | Formula of medium for culturing cordyceps spp. |
| CN102286421A (en) * | 2011-09-16 | 2011-12-21 | 中国林业科学研究院森林生态环境与保护研究所 | Liquid fermentation culture method for paecilomyces lilacinus |
| CN102293220A (en) * | 2011-09-20 | 2011-12-28 | 中国农业科学院农业环境与可持续发展研究所 | Nanocrystallized synergistic rodenticide |
| CN102293220B (en) * | 2011-09-20 | 2016-04-20 | 中国农业科学院农业环境与可持续发展研究所 | Nanocrystallized synergistic rodenticide |
| CN102382775A (en) * | 2011-11-18 | 2012-03-21 | 中国科学院微生物研究所 | Eelworm killing metarhizium and application thereof |
| CN102925405A (en) * | 2012-05-29 | 2013-02-13 | 中国林业科学研究院森林生态环境与保护研究所 | Preparation method of paecilomyces lilacinus spore powder |
| CN102925405B (en) * | 2012-05-29 | 2014-11-26 | 中国林业科学研究院森林生态环境与保护研究所 | Preparation method of paecilomyces lilacinus spore powder |
| CN102742607B (en) * | 2012-07-23 | 2014-05-07 | 中国农业大学 | Paecilomyces lilacinus fungal spore oil suspending agent and preparation method as well as application thereof |
| CN102742607A (en) * | 2012-07-23 | 2012-10-24 | 中国农业大学 | Paecilomyces lilacinus fungal spore oil suspending agent and preparation method as well as application thereof |
| CN102851219A (en) * | 2012-09-25 | 2013-01-02 | 中国热带农业科学院环境与植物保护研究所 | Paecilomyces lilacinus and application thereof |
| CN103589649B (en) * | 2013-11-08 | 2015-05-20 | 湖北森源生态科技股份有限公司 | Production method of agricultural microbial paecilomyces lilacinus for nematode prevention and control |
| CN103589649A (en) * | 2013-11-08 | 2014-02-19 | 湖北森源生态科技股份有限公司 | Production method of agricultural microbial paecilomyces lilacinus for nematode prevention and control |
| CN103749547A (en) * | 2013-12-25 | 2014-04-30 | 浙江农林大学 | Entomophthorales water floating type granules and application thereof |
| CN103749547B (en) * | 2013-12-25 | 2017-09-08 | 浙江农林大学 | A kind of entomophthora water floats type granule and application thereof |
| CN106879607A (en) * | 2015-12-15 | 2017-06-23 | 内蒙古农业大学 | Prevent and treat the biological pesticide of apple tree canker |
| CN106376602A (en) * | 2016-08-31 | 2017-02-08 | 江西天人生态股份有限公司 | Paecilomyces lilacinus granule and preparation method thereof |
| CN107586737A (en) * | 2017-09-18 | 2018-01-16 | 华中农业大学 | The Bei Laisi bacillus of one plant of anti-yellowing cucurbit wilt and its Microencapsulated Slow microbial inoculum |
| CN108294050A (en) * | 2018-03-16 | 2018-07-20 | 佛山市普尔玛农化有限公司 | Including Paecilomyces lilacinus kills line composition |
| CN110878257A (en) * | 2019-10-09 | 2020-03-13 | 北京四纬生物科技有限公司 | Paecilomyces lilacinus culture method and application |
| CN112111480A (en) * | 2020-10-14 | 2020-12-22 | 南宁汉和生物科技股份有限公司 | Method for prolonging shelf life of paecilomyces lilacinus by using double-layer embedding technology |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN101418264B (en) | Paecilomyces lilacinus and application | |
| CN101100646A (en) | Paecilomyces lilacinus and application thereof | |
| EP0706321B1 (en) | Use of streptomyces wyec 108 to control plant pathogens | |
| CN101338279B (en) | Hyphomycetes mite-killing preparation | |
| CN103184162B (en) | Trichoderma asperellum and applications thereof | |
| CN101603009B (en) | Biological fungi-proofing coniothyrium minitans ZS-1SB for preventing and treating sclerotinia as well as preparation method and application thereof | |
| CN102388922B (en) | Preparation method and application of wettable powder for preventing and controlling Sphaerotheca fuliginea trichoderma of greenhouse cucumbers | |
| CN104894035B (en) | A kind of screening technique of bacillus and its application | |
| CN113249242B (en) | Paenibacillus polymyxa and application thereof in prevention and treatment of various soil-borne diseases | |
| CN106497831A (en) | A kind of preventing and treating Phytophthora nicotianae disease composite bacteria agent capable and its preparation method and application | |
| CN104222076A (en) | Bacillus methylotrophicus wettable powder and preparation method and application thereof | |
| CN101352177B (en) | Compound successive crop-resistance micro-ecological formulation special for cotton and special bacterial strain and use thereof | |
| CN102021122B (en) | High-efficiency insecticidal fungus and applications thereof | |
| CN107254317A (en) | A kind of preprocess method of stalk and the renovation agent for including stalk after pretreatment | |
| CN101990907A (en) | Composite biological antimicrobial for preventing wilt and greensickness of cotton | |
| CN105039184A (en) | Beauveria bassiana strain having pathogenicity on cyclophragma undans and application thereof | |
| CN117448192A (en) | Bacillus bailii XU183 and application thereof | |
| CN104531571B (en) | Pseudomonas fluorescens and biological preparation and application in preventing and controlling sugarcane smut | |
| CN103451112B (en) | Saline-alkali tolerant trichoderma longibrachiatum and application thereof | |
| CN103451111B (en) | Saline-alkali tolerant trichoderma harzianum and application thereof | |
| CN105154339A (en) | Trichoderma viride strain and application thereof | |
| CN105132296A (en) | Hook-like trichoderma strain and application thereof | |
| CN102408996B (en) | Wettable powder of biocontrol bacteria coniothyrium minitans Chy-1C-1 and application | |
| CN103392744A (en) | Trichoderma preparation for controlling pepper phytophthora blight and field tank mix pesticide containing the same | |
| CN102960369B (en) | Preparation method of granular paecilomyces lilacinus biological nematocide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C02 | Deemed withdrawal of patent application after publication (patent law 2001) | ||
| WD01 | Invention patent application deemed withdrawn after publication |