CN102206587A - Culture method of sclerotium of beauveria bassiana and application thereof - Google Patents
Culture method of sclerotium of beauveria bassiana and application thereof Download PDFInfo
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Abstract
Aiming at the defects of conidiospores as propagula of an insecticide of insect pathogenic fungi, the invention provides a technology of liquid culture of entomogenous fungus sclerotia, and the entomogenous fungus can be beauveria bassiana. The liquid culture method of entomogenous fungus sclerotia provided by the invention is required to control the liquid culture components and the culture conditions within a certain range. The cultured sclerotia can be stored for a long time after vacuum drying, and can still germinate mycelia and pathogenic conidiospores under suitable conditions. The micro-sclerotia prepared by drying have maintained activity, can germinate mycelia and spores after rehydration, and increase the potential of beauveria bassiana as a biocontrol factor for western flower thrips in a soil-dwelling phase. The technology has low cost, long storage period of sclerotia, and is quite suitable for popularization.
Description
Technical field
The invention belongs to biological technical field, be specifically related to the cultural method and the application aspect pest control thereof of entomogenous fungi sclerotium.
Background technology
Frankliniella occidentalis Frankliniella occidentalis (Pergande) is a kind of important pests on the global diversified economy crop (Thysanoptera:Thripidae).Frankliniella occidentalis causes seriously crop by file absorption food and propagation plant virus causes harm.Comprise the life history of Frankliniella occidentalis and getting the food stage on the ground (adult, 1 age and 2 age nymph) and soil dwell the stage (2 nymph latter stages in age, prepupa and pupa).Most of Frankliniella occidentalis aging 2 age nymph all show positive geotaxis and negative phototaxis, transfer to soil or the potted plant medium from plant and pupate.According to the difference of host plant kind, there is 98% thrips to pupate at most, and is distributed in the dark soil layer of 1.5～2.0cm underground, cause harm up to growing for from soil, sprouting wings to come out to transfer to again on the ground behind the adult.In addition, owing to use chemical agent continually repeatedly, this worm has been developed the population that number of chemical sterilant tool resistance.
Use entomogenous fungi in the soil and be and effectively administer the dwell effective measure in stage of thrips soil.Beauveria bassiana (Beauveria bassiana (Balsamo) Vuillemin is hereinafter to be referred as muscardine) is that the host composes one of important insect pathogenic fungus widely, and the muscardine bacterial strain of many commercializations or non-commercialization can cause that all the height of Frankliniella occidentalis infects.Promoted the development of corresponding formulation with the commercial interest of entomogenous fungi control subterranean pest-insect.The basis of these formulations is that liquid phase is cultivated mycelia that produces or the conidium that the solid phase cultivation produces on nutrition or non-nutrition carrier.These mycelia or conidium normally drought tolerance difference and shelf-life are shorter, thereby have limited their practical application.
In order to survive in soil and putrid vegetable material, many plant pathogenic fungis produce sclerotium, i.e. mycelia aggregate closely, and with these propaguluies as the structure of surviving the winter.Some plant pathogenic fungis, for example the brown red downright bad germ Mycoleptodiscus terrestris of soybean anthracnose bacterium Colletotrichum truncatum (Schweinitz) and soybean (Gerd.) is in the deep-layer liquid cultivation and fermentation, can produce germ nuclear (microsclerotia, MS), i.e. the sclerotium particle of diameter 200～600 μ m.As mycoherbicide, the MS of these weeds pathogens holds in soil and the aquatic environment to imitate infectious propagulum.Yet up to the present also control Frankliniella occidentalis soil not being dwelt, important pathogenic fungi of stage-muscardine MS cultivates and the report of application.With insects such as insect pathogenic fungus sclerotium control Frankliniella occidentalis, can simplify the production technique of pest-resistant preparation, reduce cost, and environment is not polluted, therefore, have broad application prospects.
Summary of the invention
At the defective of conidium as the propagulum existence of insect pathogenic fungus sterilant, the invention provides a kind of technology of liquid culture entomogenous fungi sclerotium, described entomogenous fungi is preferably beauveria bassiana.The liquid cultivating method of entomogenous fungi sclerotium provided by the present invention need be controlled at liquid culture composition and culture condition in certain scope.The sclerotium of turning out is through after the vacuum-drying, can Long-term Storage, and under suitable condition, still can sprout mycelia and the conidium of virulence is arranged, this technical costs is low, and sclerotium storage period is long, is fit to very much popularize.
Purpose of the present invention realizes by following technical proposals:
A kind of cultural method of entomogenous fungi sclerotium is characterized in that may further comprise the steps: 1) one-level is dull and stereotyped cultivates; 2) preparation of spore suspension; 3) liquid culture sclerotium; 4) drying of sclerotium.
Described entomogenous fungi comprises any insect pathogenic fungus, is preferably beauveria bassiana.
Described one-level is dull and stereotyped cultivates is that fungal bacterial strain is cultivated on the PDA substratum and treated that it produces spore, and with the spore-bearing agar block that is cut into, the very low temperature preservation is standby in glycerine.Can under the fungus culture condition of routine, carry out during cultivation, as: cultivated for 3 weeks on the PDA substratum in (～22 ℃) under the room temperature, spore-bearing agar plate is cut into 1mm
2Agar block, be immersed in 10% glycerine in-80 ℃ and preserve down.
The preparation of described spore suspension is that the agar block with chilled storage is inoculated on the PDA flat board, cultivated for 2～3 weeks down in room temperature (～22 ℃), add the sterilized water contain 0.1% polyoxyethylene glycol oleic acid based compound (tween-80), mixing, dilution are broken up in aseptic technique, and (final concentration is 1.0 * 10 to make spore suspension
9L
-1) standby.
Described liquid culture sclerotium is with step 2) spore suspension of preparation inserts in the nutrient solution and (makes the spore final concentration reach 5 * 10
6L
-1), be positioned in the constant temperature shaking table and cultivate.Culture condition is: temperature 22-28 ℃, shake fast 280-300rpm; Be preferably 25 ℃ of culture temperature, shake fast 300rpm.
The drying of described sclerotium is to add diatomite (DE) in the nutrient solution with step 3), make concentration reach the 5gDE/100mL nutrient solution, with MS-DE mixture suction filtration to preweighted filter paper, separate from filter paper, with freeze drier vacuum-drying to water content＜5% o'clock, freezing sealing, and in 4 ℃ of storages.
PDA substratum used in the above-mentioned sclerotium culturing process is microbiological culture media commonly used, and its composition and content all are known, can check in by textbook or laboratory manual.
Nutrient solution composition used during the liquid culture sclerotium comprises: basic salt (gL
-1), trace element (μ gL
-1), vitamin B complexes (μ gL
-1), nitrogenous source (gL
-1), carbon source (gL
-1) and tween-80 (v/v); Wherein, described basic salt (gL
-1) be selected from: KH
2PO
40.5-2.0, MgSO
47H
2O 0.5-2.5 or CaCl
22H
2O 0.01-0.05; Described trace element (μ gL
-1) be selected from: H
3BO
35-30, MnSO
44H
2O 10-50, CuSO
45H
2O 50-80, ZnSO
47H
2O 50-400 or FeCl
36H
2O 20-500; Described vitamin B complexes (μ gL
-1) be selected from: thiamines 5-30, Riboflavin Tetrabutyrate-15, Pyridoxylamine 0.5-2 or dextro calcium pantothenate 2-10; Described nitrogenous source (gL
-1) be selected from: soyflour 5-15 or Semen Maydis powder, 5-15; Described carbon source (gL
-1) be glucose, 5-25; Described tween-80 (v/v) is 0.2-0.8%; The final pH of nutrient solution transfers to 5.5-7.1.Be preferably: wherein said basic salt (gL
-1) be selected from: KH
2PO
41.5, MgSO
47H
2O 0.5 or CaCl
22H
2O 0.01; Described trace element (μ gL
-1) be selected from: H
3BO
330, MnSO
44H
2O 40, CuSO
45H
2O 80, ZnSO
47H
2O400 or FeCl
36H
2O 500; Described vitamin B complexes (μ gL
-1) be selected from: thiamines 30, riboflavin 15, Pyridoxylamine 2 or dextro calcium pantothenate 10; Described nitrogenous source (gL
-1) be selected from soyflour 15 or Semen Maydis powder 15; Described carbon source (gL
-1) be glucose 25; Described tween-80 (v/v) is 0.4-0.8%; The final pH of nutrient solution transfers to 6.8.
The invention still further relates to the application of entomogenous fungi sclerotium in the biological prevention and control agent of preparation pest control that obtains by above-mentioned cultural method, described entomogenous fungi preferred spheres beauveria bassiana, the preferred Frankliniella occidentalis of described insect.
The present invention also further relates to a kind of biological prevention and control agent of pest control, it is characterized in that, comprising: by the entomogenous fungi sclerotium and the auxiliary material of above-mentioned cultural method acquisition.Described entomogenous fungi sclerotium preferred spheres beauveria bassiana sclerotium, the preferred Frankliniella occidentalis of described insect; Described auxiliary material is a pest-resistant formulation preparation field auxiliary reagent commonly used, and those skilled in the art fully can be according to the type selecting corresponding auxiliary material type of preparation.
Fungal bacterial strain that the present invention is used and worm source as beauveria bassiana and Frankliniella occidentalis, all belong to public experiment material, can obtain by conventional approach, as the commercial sources purchase etc.For example, among the embodiment used beauveria bassiana available from INST OF AGRICULTURAL RESOURCES.
The present invention can produce the entomogenous fungi sclerotium with the liquid submerged fermentation method first.The germ nuclear energy of dry preparation enough maintains vigour, and can sprout mycelia and spore after the rehydration, has increased above-mentioned fungi prevents and treats the factor as the Frankliniella occidentalis soil stage of dwelling potentiality.This production process technology is simple, and is with low cost, and the nonstaining property waste material produces in the production process, very environmental protection.Insects such as entomogenous fungi control Frankliniella occidentalis with this method obtains can reduce the pollution of chemical pesticide to plant and environment, and to not injury of humans and animals, therefore, the present invention has broad application prospects.
Description of drawings:
The growth of beauveria bassiana sclerotium when Fig. 1 deep liquid on shaking table is cultivated.
The conidium of inserting after (A) is 24 hours begins to form the mycelia of even matter.Along with the increase of incubation time, mycelia begins to assemble the formation sclerotium; (B) Day 2; (C) Day 4; (D) Day 6.
Embodiment
Fungal bacterial strain that the present invention is used and worm source as beauveria bassiana and Frankliniella occidentalis, all belong to public experiment material, can obtain by conventional approach, as the commercial sources purchase etc.For example, among the embodiment used beauveria bassiana available from INST OF AGRICULTURAL RESOURCES.
The cultivation of embodiment 1 beauveria bassiana sclerotium:
1. cultural method:
1.1 the dull and stereotyped spawn culture of one-level
The beauveria bassiana bacterial strain was cultivated for 3 weeks on the PDA substratum in (～22 ℃) under the room temperature.Spore-bearing agar plate is cut into 1mm
2Agar block, be immersed in 10% glycerine in-80 ℃ and preserve down.
1.2 the preparation of spore suspension
During liquid culture, the agar block of chilled storage is inoculated on the PDA flat board, cultivated for 2～3 weeks down in room temperature (～22 ℃).Add and to contain 0.1% polyoxyethylene glycol oleic acid based compound (mixing, dilution are broken up in aseptic technique for tween-80, sterilized water Sigma), and making final concentration is 1.0 * 10
9L
-1Spore suspension standby.
1.3 liquid culture sclerotium
In the 250mL beaker, the 100mL nutrient solution of packing into inserts spore suspension, makes the spore final concentration reach 5 * 10
6L
-1, be positioned over constant temperature shaking table (25 ℃ of temperature; Shake fast 300rev.min
-1) the middle cultivation.Through hand rolling beaker commonly used, to reduce the growth of mycelia on walls of beaker.And after inoculation the form of resultant in the microscopic examination nutrient solution day by day, detection of biological amount accumulation in the time of the 6th day, spore concentration and MS concentration.In each experiment, from each beaker, get sample 2 times, every kind of nutrient solution is provided with 3 beakers and repeats.All experiments repeat 2 times at least.
Described nutrient solution composition comprises: (1) basic salt (gL
-1): KH
2PO
4, 1.5, MgSO
47H
2O, 0.5, CaCl
22H
2O, 0.01; (2) trace element (μ gL
-1): H
3BO
3, 30, MnSO
44H
2O, 40, CuSO
45H
2O, 80g; ZnSO
47H
2O, 400, FeCl
36H
2O, 500; (3) vitamin B complexes (μ gL
-1): thiamines, 30, riboflavin, 15, Pyridoxylamine, 2, dextro calcium pantothenate, 10; (4) nitrogenous source (gL
-1): soyflour, 15, Semen Maydis powder, 15; (5) carbon source (gL
-1): glucose, 25; (6) tween-80 (v/v): 0%, 0.1%, 0.2%, 0.4%, 0.6%, 0.8%, 1.0% or 1.2%.The final pH of nutrient solution transfers to 6.8.Keep in test basic salt, trace element, vitamin B complexes, carbon source to fix, detect the result who adds different sorts nitrogenous source and different concns tween 80 combination back gained.
When detecting the cumulative biomass, get the 1mL nutrient solution from cultivate beaker, vacuum filtration is to preweighted filter paper.Filter paper under room temperature vacuum-drying to stable weight.Change calculations cumulative biomass according to weight before and after the filter paper.When detecting the germ nuclear concentration, at first nutrient solution is diluted to suitable MS counting, gets 100 μ L nutrient solutions then on glass slide, covered for degree.All MS on the microscopic count slide.Have only diameter to surpass 50 μ m, independently assemble mycelium and just count MS.When nutrient solution dilution and sampling, continue to rock to guarantee evenly.When detecting spore concentration, at first nutrient solution is diluted to suitable spore counting and is degree.Get 100 μ L nutrient solutions then on blood counting chamber, covered.Spore count is calculated in 5 samplings.
1.4 the drying of sclerotium
Beauveria bassiana adds diatomite (DE) after cultivating 8d in nutrient solution, make concentration reach the 5gDE/100mL nutrient solution.With recirculated water vacuum pump (SHZ-III, Shanghai Yarong Biochemical Instrument Plant) with MS-DE mixture suction filtration to preweighted Whatman No.54 filter paper.The MS-DE mixture is separated from filter paper, with freeze drier (LGJ-10D, Fourth Ring, Beijing scientific instrument company limited of instrument plant) vacuum-drying is to water content＜5% o'clock, and (the sharp light industry and machinery of Shanghai ancient cooking vessel Manufacturing Co., Ltd) is packaged in the aluminium foil bag with sealing machine, and in 4 ℃ of storages.
2. nitrogenous source is to the influence of beauveria bassiana germ nuclear, gemma and biomass:
Initial fermenting experiment shows (table 1), at fixed base salt, vitamin B complexes, trace element is when carbon source and 0.4% tween 80 are constant, after inserting the beauveria bassiana spore suspension and cultivating 5d, with soyflour and Semen Maydis powder nutrient solution as nitrogenous source, it is smooth to observe the edge, well-developed sclerotium, and there is significant difference (P＜0.05) in quantity between the two.
Table 1 nitrogenous source is to the influence of beauveria bassiana biomass, gemma and germ nuclear output
Annotate: the different letter representations of going together are in 0.05 level difference remarkable (P＜005).
3. beauveria bassiana germ karyogenesis pattern:
It with the Semen Maydis powder nitrogenous source, contain in the nutrient solution of 0.4% tween-80, such pattern (Fig. 1) is followed in the formation of culture behind the inoculation beauveria bassiana spore suspension: conidium sprouts the mycelia that forms even matter in 24h, the 2nd day, mycelial density increased.Through after the 2d growth, mycelia begins to assemble the formation sclerotium, during at last by the 6th day, forms entire sclerotium again.The beauveria bassiana germ nuclear that forms varies in size, and diameter range is 50～300 μ m.
4. tween 80 is to the influence of beauveria bassiana germ nuclear and gemma output: the tween-80 that adds different content in the nutrient solution that with the Semen Maydis powder is nitrogenous source is examined by the germ of beauveria bassiana and the formation of gemma has remarkably influenced (table 2).Tween-80 content is at 0.4%～0.8% o'clock in the nutrient solution, can form physically well develop, the even germ nuclear of (diameter 50～300 μ m) of size; When the content of tween-80 in the nutrient solution≤0.2%, except the germ nuclear that forms diameter 50～300 μ m, also form some bigger germs nuclears (500～1000 μ m), and germ nuclear volume significantly descend (P＜0.05); When the content of tween-80 in the nutrient solution was 1.0%～1.2%, hyphal fragment or gemma quantity significantly increased (P＜0.05), but did not observe the formation of germ nuclear.
Table 2 tween-80 to beauveria bassiana germ nuclear output, the germ nucleome is long-pending and the influence of gemma output
Annotate: the different letter representations of going together are in 0.05 level difference remarkable (P＜005).
Embodiment 2 gives birth to test and tests:
1. give birth to the detection of surveying efficient:
The raising of two flower thripss: in the cylinder glass container of 0.5L, put into 3～4 beanpods, the thrips adult transferred in the container lay eggs, container is placed in the growth case, culture condition is set to 26 ℃ of temperature, relative humidity 60%～70%, photoperiod is L: D=13: 11, remove adult behind the 12h of laying eggs to guarantee the unanimity of instar larvae.The larva (L2) of collection 2d in 2 age is standby behind the 8d.
With vermiculite, clay loam and nutrition soil mixed by 2: 1: 1.The MS-DE uniform particles is joined in the soil that is mixed, and mixture ratio is 100 μ g/100g soil.Said mixture is packed in the plastic flowerpot, and evenly spray deionized water, till having moisture content to ooze out at the bottom of the basin.Flowerpot is put into plastic pallet, often add water and keep continuing that water is arranged in the pallet, and can be siphoned in the plastic tub.Control soil is handled equally, but does not add the MS-DE particle.
After sclerotium is mixed into 7d in the soil (with abundant product spore), with the fine, soft fur brush 30 L2 thrips larvas are transferred to (as food) on a bit of beanpod, beanpod is put into the surface of above-mentioned dress basin soil.The cylindrical plastic container of the both ends open that button is transparent on the basin, the plastic containers upper end is covered with anti-thrips nylon yarn (120 order).Be added with the adult of the yellow insect-sticking plate of a 5cm * 4cm with trapping emergence in plastic containers inwall upper end.The plastic tub that is covered with plastic containers is placed in the growth case cultivates, and culture condition is provided with the same.Each is handled and repeats 5 times, and whole experiment repeats 2 times.
After inserting L2 larva 4d, the thrips adult begins to sprout wings, and adheres on the interior insect-sticking plate of plastic containers, and perhaps at soil surface, perhaps at the inner edge of plastic containers, observe adult eclosion with magnifying glass every day, and 7d is up to thrips adult appearance never again continuously.Whether the pupa and the adult of collecting from soil surface or plastic containers inner edge all are put on the insect-sticking plate, be placed on together with the insect-sticking plate that lures adult to cultivate (25 ℃) in the culture dish that is lined with wet filter paper and infected deadly by beauveria bassiana to detect them.Statistics is by lethal bombys batryticatus of fungi infestation and healthy adult.
2. statistical study: the one-tenth borer population that with sprouting wings in the soil that contrasts deducts the emergence that obtains in the soil of processing and becomes borer population, obtains thrips accumulation death toll.The mortality ratio of gained is carried out the sine conversion.Biomass in the nutrient solution, gemma and MS output, the average number average of exsiccant MS-DE particulate conidium output and thrips mortality ratio adopts the unitary variant variance analysis---least significant difference test (SPSS 10.0, One-Way ANOVA:LSD tests).
3. beauveria bassiana MS-DE particle is to the dwell preventive effect in stage of Frankliniella occidentalis soil:
The Frankliniella occidentalis soil corrected mortality (mean number ± standard error) in stage of dwelling in the soil that table 3 is handled with beauveria bassiana germ nuclear
Annotate: the different letter representations of going together are remarkable at 0.05 level difference
As can be seen from Table 3, compare with untreated control, the muscardine germ nuclear premix that no matter to be soyflour or Semen Maydis powder turn out as nitrogenous source can both significantly improve the dwell mortality ratio (P＜0.05) in stage of Frankliniella occidentalis soil in the soil.
Above-mentioned having experimental results show that with the liquid submerged fermentation method can produce muscardine germ nuclear.The germ nuclear energy of dry preparation enough maintains vigour, can sprout mycelia and spore after the rehydration, has increased muscardine as the dwell potentiality of the stage biological and ecological methods to prevent plant disease, pests, and erosion factor of Frankliniella occidentalis soil.
Claims (13)
1. the cultural method of an entomogenous fungi sclerotium is characterized in that may further comprise the steps: 1) the dull and stereotyped cultivation of one-level; 2) preparation of spore suspension; 3) liquid culture sclerotium; 4) drying of sclerotium.
2. cultural method according to claim 1 is characterized in that: described entomogenous fungi is a beauveria bassiana.
3. cultural method according to claim 1 and 2 is characterized in that: the one-level of described step 1) is dull and stereotyped cultivates is that fungal bacterial strain is cultivated on the PDA substratum and treated that it produces spore, and with the spore-bearing agar block that is cut into, the very low temperature preservation is standby in glycerine.
4. according to the described cultural method of one of claim 1-3, it is characterized in that: the preparation of spore suspension described step 2) is that the agar block with chilled storage is inoculated on the PDA flat board, at room temperature cultivated for 2～3 weeks, add the sterilized water contain 0.1% polyoxyethylene glycol oleic acid based compound, mixing, dilution are broken up in aseptic technique, and it is standby to make spore suspension.
5. according to the described cultural method of one of claim 1-4, it is characterized in that: the liquid culture sclerotium of described step 3) is with step 2) spore suspension of preparation inserts in the nutrient solution, be positioned in the constant temperature shaking table and cultivate, culture condition is: temperature 22-28 ℃, shake fast 280-300rpm, 25 ℃ of preferred culture temperature are shaken fast 300rpm.
6. according to the described cultural method of one of claim 1-5, it is characterized in that: used nutrient solution is formed and is comprised during described step 3) liquid culture sclerotium: basic salt (gL
-1), trace element (μ gL
-1), vitamin B complexes (μ gL
-1), nitrogenous source (gL
-1), carbon source (gL
-1) and tween-80 (v/v); Wherein, described basic salt (gL
-1) be selected from: KH
2PO
40.5-2.0, MgSO
47H
2O 0.5-2.5 or CaCl
22H
2O 0.01-0.05; Described trace element (μ gL
-1) be selected from: H
3BO
35-30, MnSO
44H
2O 10-50, CuSO
45H
2O 50-80, ZnSO
47H
2O 50-400 or FeCl
36H
2O 20-500; Described vitamin B complexes (μ gL
-1) be selected from: thiamines 5-30, Riboflavin Tetrabutyrate-15, Pyridoxylamine 0.5-2 or dextro calcium pantothenate 2-10; Described nitrogenous source (gL
-1) be selected from: soyflour 5-15 or Semen Maydis powder, 5-15; Described carbon source (gL
-1) be glucose 5-25; Described tween-80 (v/v) is 0.2-0.8%; The final pH of nutrient solution transfers to 5.5-7.1.
7. cultural method according to claim 6 is characterized in that: used nutrient solution is formed and is comprised during described step 3) liquid culture sclerotium: basic salt (gL
-1), trace element (μ gL
-1), vitamin B complexes (μ gL
-1), nitrogenous source (gL
-1), carbon source (gL
-1) and tween-80 (v/v); Wherein said basic salt (gL
-1) be selected from: KH
2PO
41.5, MgSO
47H
2O0.5 or CaCl
22H
2O 0.01; Described trace element (μ gL
-1) be selected from: H
3BO
330, MnSO
44H
2O 40, CuSO
45H
2O 80, ZnSO
47H
2O 400 or FeCl
36H
2O 500; Described vitamin B complexes (μ gL
-1) be selected from: thiamines 30, riboflavin 15, Pyridoxylamine 2 or dextro calcium pantothenate 10; Described nitrogenous source (gL
-1) be selected from soyflour 15 or Semen Maydis powder 15; Described carbon source (gL
-1) be glucose 25; Described tween-80 (v/v) is 0.4-0.8%; The final pH of nutrient solution transfers to 6.8.
8. the application of entomogenous fungi sclerotium in the biological prevention and control agent of preparation pest control that obtains with the described cultural method of one of above-mentioned claim 1-7.
9. application according to claim 8 is characterized in that: described entomogenous fungi is a beauveria bassiana.
10. according to Claim 8 or 9 described application, it is characterized in that: described insect is a Frankliniella occidentalis.
11. the biological prevention and control agent of a pest control is characterized in that, comprising: the entomogenous fungi sclerotium and the auxiliary material that require the described cultural method acquisition of one of 1-7 by aforesaid right.
12. biological prevention and control agent according to claim 11 is characterized in that, described entomogenous fungi sclerotium is a beauveria bassiana.
13., it is characterized in that described insect is a Frankliniella occidentalis according to claim 11 or 12 described biological prevention and control agents.
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Cited By (5)
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| CN106497805A (en) * | 2016-12-30 | 2017-03-15 | 四川省农业科学院蚕业研究所 | A kind of cultural method of beauveria bassiana |
| CN108949588A (en) * | 2018-08-23 | 2018-12-07 | 重庆大学 | A kind of application of the preparation method and preparation of beauveria bassiana Microsclerotia and its preparation |
| US11533916B2 (en) * | 2019-03-08 | 2022-12-27 | Ypf Tecnología S.A. | Method of preparation of microsclerotia of Beauveria bassiana |
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| 张素华，等: "不同温度下4株白僵菌对西花蓟马的致病力", 《植物保护》 * |
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| CN104212724B (en) * | 2014-09-01 | 2017-04-19 | 中国农业科学院植物保护研究所 | Beaueria bassaria (Balsamo) Vuillemin bacterial strain GZGY-1-3 and application thereof |
| CN106497805A (en) * | 2016-12-30 | 2017-03-15 | 四川省农业科学院蚕业研究所 | A kind of cultural method of beauveria bassiana |
| CN108949588A (en) * | 2018-08-23 | 2018-12-07 | 重庆大学 | A kind of application of the preparation method and preparation of beauveria bassiana Microsclerotia and its preparation |
| WO2020038020A1 (en) * | 2018-08-23 | 2020-02-27 | 重庆大学 | Microsclerotia of beauveria bassiana, preparation method for formulation of same, and use of formulation |
| CN108949588B (en) * | 2018-08-23 | 2020-07-28 | 重庆大学 | Preparation method of beauveria bassiana microsclerotia and preparation thereof and application of preparation |
| EP3744827A4 (en) * | 2018-08-23 | 2021-07-07 | Chongqing University | Microsclerotia of beauveria bassiana, preparation method for formulation of same, and use of formulation |
| US11219220B2 (en) | 2018-08-23 | 2022-01-11 | Chongqing University | Preparation method of Beauveria bassiana microsclerotium and formulation thereof, application of formulation thereof |
| US11533916B2 (en) * | 2019-03-08 | 2022-12-27 | Ypf Tecnología S.A. | Method of preparation of microsclerotia of Beauveria bassiana |
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