CN109757521A - Muscardine microbial inoculum and its application - Google Patents
Muscardine microbial inoculum and its application Download PDFInfo
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- CN109757521A CN109757521A CN201910109413.6A CN201910109413A CN109757521A CN 109757521 A CN109757521 A CN 109757521A CN 201910109413 A CN201910109413 A CN 201910109413A CN 109757521 A CN109757521 A CN 109757521A
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Abstract
The invention belongs to agricultural technology fields, and in particular to a kind of muscardine microbial inoculum and its application.Muscardine microbial inoculum of the invention includes muscardine BRNS50206, mixed support agent, mineral carrier and cereal carrier, wherein the deposit number of muscardine BRNS50206 is CGMCC No.15993.The present invention by muscardine BRNS50206 as microbial inoculum active component, it is obligate to infect insect to vertebrate safety, have no toxic side effect, it is lossless pollution-free with environment NATIVE FUSION, be the biological pesticide of Environmental Safety；Present invention can apply to prevent and treat in the kind class pest such as crops, forest, lawn, the grub of herbage, root weevil, cutworm.
Description
Technical field
The invention belongs to agricultural technology fields, and in particular to muscardine microbial inoculum and its application.
Background technique
Subterranean pest-insect includes more than 200 kinds of chrysomelid grub, root weevil, root, cutworm, mole cricket, root maggot etc., in life cycle
At least one stage lives in the soil, seed, seedling, rhizome and the ground of the plants such as feeding crop, forest, flowers, herbage
Lower fruit, the sprout tumble that causes to be short of seedling, plant are wilted even death because of nutrient absorption deficiency, are caused harm hidden, it is difficult to prediction.
For a long time, China depends on chemical pesticide control pest, due to chemical insecticide it is a large amount of, year after year apply
With agricultural product, soil and waters have constituted a serious threat to human health and damaged by pesticide residual contamination.The anti-medicine of pest
Property enhancing so that dosage is continuously increased, expenses for prevention and control multiplication；Largely damaged beneficial to natural enemy biology, ecological environment by
It is serious to destroy, lead to the second rice crop season of pest.For the yield, quality and safety for preserving the ecological environment, guaranteeing food and feed,
Agricultural pests are controlled using biological prevention large area, take sustainable pest management measure imperative
Insect pathogenic fungus is most important insecticidal microorganism monoid, and about 70% insect disease is true under field conditions (factors)
It is microbial.Its pathogenic mechanism is complicated, and pest will not be caused to generate resistance, to the resistance problems great potential for solving pest.
Therefore, insect pathogenic fungus is as important substitution pesticide.Wherein, muscardine (Beauveria brongniartii) is
A kind of insect pathogenic fungus, parasitic 7 70 various insects of purpose of energy penetrate polypide by being attached to insect cuticle, generating germ tube
Epidermis intrusion, consumption polypide nutrition, itself quickly breeding, generation toxin, cause body tissue damage and lesion dead.
However, Field information finds that muscardine of manual application etc. has certain Natural Attenuation in field, and it infects mesh
Mark insect needs certain probability and pathogenic course time therefore to tend not to reach ideal control efficiency.
Summary of the invention
The purpose of the present invention is to provide a kind of muscardine microbial inoculums.
A further object of the present invention is to provide the applications of above-mentioned muscardine microbial inoculum.
The muscardine microbial inoculum for being used to prevent and treat subterranean pest-insect of specific embodiment according to the present invention, including the white deadlock of Bu Shi
Bacterium BRNS50206, mixed support agent, mineral carrier and cereal carrier, wherein the deposit number of muscardine BRNS50206
For CGMCC No.15993.
The muscardine microbial inoculum for being used to prevent and treat subterranean pest-insect of specific embodiment according to the present invention comprising following weight
Measure part raw material: 5~50 parts of muscardine conidia powder, 4~10 parts of mixed support agent, 20~60 parts of mineral carrier, cereal carrier
10~70 parts, wherein hundred million/g of content range 25-250 of muscardine conidia powder.
The muscardine microbial inoculum of specific embodiment according to the present invention, including following raw material: muscardine
15~25 parts of mycelium, 4~10 parts of mixed support agent, 20~60 parts of mineral carrier, 10~70 parts of cereal carrier, wherein mycelia
Body is pressed to be calculated containing dry weight 2.5%.
The muscardine microbial inoculum of specific embodiment according to the present invention, mixed support agent be dialkyl succinylsuccinate sodium sulfonate,
Hemicellulose, sodium alginate, chitosan are mixed to get according to mass ratio for 15:5:4:1.
The muscardine microbial inoculum of specific embodiment according to the present invention, which is characterized in that the mineral carrier is bumps
One of stick soil, talcum powder, diatomite, white carbon black, calcium carbonate, clay are a variety of.
The muscardine microbial inoculum of specific embodiment according to the present invention, the cereal carrier are peanut pod shell powder, corn
One of core powder, corn stalk powder, wheat straw powder, rice straw powder, sugar beet powder, bagasse powder are a variety of.
The muscardine microbial inoculum of specific embodiment according to the present invention, the muscardine microbial inoculum further includes adhesive
0.1~6 part, described adhesive is polyvinyl alcohol or carboxymethyl cellulose.
The muscardine microbial inoculum of specific embodiment according to the present invention, the muscardine microbial inoculum further includes synergist
1~4 part.
The muscardine microbial inoculum of specific embodiment according to the present invention, the synergist are 10% imidacloprid wettable powder
Agent, 25% Diacloden wettable powder or 20% Rynaxypyr suspending agent.
Beneficial effects of the present invention:
The present invention by muscardine BRNS50206 as microbial inoculum active component, it is obligate to infect insect and to vertebrate
Safety, has no toxic side effect, lossless pollution-free with environment NATIVE FUSION, is the biological pesticide of Environmental Safety；Present invention can apply to
It prevents and treats in the kind class pest such as crops, forest, lawn, the grub of herbage, root weevil, cutworm.
The bioactivity and field survival ability of mixed support agent protection muscardine BRNS50206；
Synergist and muscardine generate synergistic effect, accelerate infecting for muscardine, improve infect efficiency；
Mineral carrier, cereal carrier also have auxiliary protection muscardine while load and divided active component
Activity, chesson environment, the effect for promoting the survival of muscardine field and infecting；
Adhesive can be added when manufacturing granule, spread application convenient for forming, conducive to throwing.
Detailed description of the invention
Fig. 1 shows microbial inoculum of the present invention to the toxicity test result of black cutworm.
The deposit number of muscardine (Beauveria brongniartii) BRNS50206 bacterial strain of the invention is
CGMCC No.15993, the bacterial strain were stored in China General Microbiological Culture Collection Center on 07 17th, 2018, in preservation
Heart location: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica.
Specific embodiment
Embodiment 1
(1) by strain spore inoculating to fluid nutrient medium, wherein containing sucrose 3%, peptone 1%, yeast extract powder
0.25%, magnesium sulfate 0.01%, surplus are water, cultivate 48-60 hours, obtain liquid seeds；Then liquid culture is inoculated with
To solid medium, wherein leather shell containing cereal 35~60%, wheat bran 35~60%, corn flour 5~10%, after culture 5~9 days,
Matrix is directly crushed after drying or extraction obtains conidia powder；
(2) by weight preparation dialkyl succinylsuccinate sodium sulfonate: hemicellulose: sodium alginate: chitosan=15:5:4:1
Mixed support agent；
(3) 5 parts by weight conidia powders, 4 parts by weight adjuvants are mixed with 10 parts by weight peanut shell powders；
(4) 1 parts by weight, 10% imidacloprid wettable powder is added in 20 parts by weight attapulgites, is uniformly mixed；
(5) mixture that step (3), (4) obtain is mixed again, obtains muscardine fine granule.
Embodiment 2
(1) by strain spore inoculating to fluid nutrient medium, wherein containing sucrose 3%, peptone 1%, yeast extract powder
0.25%, magnesium sulfate 0.01%, surplus are water, cultivate 48-60 hours, obtain liquid seeds；Then liquid culture is inoculated with
To solid medium, wherein leather shell containing cereal 35~60%, wheat bran 35~60%, corn flour 5~10%, after culture 5~9 days,
Matrix is directly crushed after drying or extraction obtains conidia powder；
(2) by weight preparation dialkyl succinylsuccinate sodium sulfonate: hemicellulose: sodium alginate: chitosan=15:5:4:1
Mixed support agent；
(3) 20 parts by weight conidia powders, 6 parts by weight adjuvants are mixed with 10 parts by weight Corn core powder；
(4) 2 parts by weight, 25% Diacloden wettable powder is added in 20 parts by weight diatomite, is uniformly mixed；
(5) mixture that step (3), (4) obtain is mixed again, obtains muscardine fine granule.
Embodiment 3
(1) by strain spore inoculating to fluid nutrient medium, wherein containing sucrose 3%, peptone 1%, yeast extract powder
0.25%, magnesium sulfate 0.01%, surplus are water, cultivate 48-60 hours, obtain liquid seeds；Then liquid culture is inoculated with
To solid medium, wherein leather shell containing cereal 35~60%, wheat bran 35~60%, corn flour 5~10%, after culture 5~9 days,
Matrix is directly crushed after drying or extraction obtains conidia powder；
(2) by weight preparation dialkyl succinylsuccinate sodium sulfonate: hemicellulose: sodium alginate: chitosan=15:5:4:1
Mixed support agent；
(3) 50 parts by weight conidia powders, 10 parts by weight adjuvants are mixed with 70 parts by weight bagasse powder；
(4) 3 parts by weight, 10% imidacloprid wettable powder is added in 60 parts by weight attapulgites, is mixed；
(5) it finally by the mixture of 6 parts by weight carboxymethyl celluloses and above-mentioned steps (3) and step (4), remixes
Even, extrusion granulation obtains muscardine granule.
Embodiment 4
(1) by strain spore inoculating to level liquid culture medium, wherein containing sucrose 3%, peptone 1%, yeast extract powder
0.25%, magnesium sulfate 0.01%, surplus are water, cultivate 48-60 hours, obtain level liquid object；Then it transfers again as strain
To culture medium containing secondary liquid, wherein containing sucrose 2%, soluble starch 1%, brewer's yeast 1%, carrot dry powder 0.4-
0.6%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.05%, surplus are water, cultivate 48-72 hours, obtain secondary liquid culture；
Most of moisture is removed by centrifugation, obtains concentration mycelium；
(2) by weight preparation dialkyl succinylsuccinate sodium sulfonate: hemicellulose: sodium alginate: chitosan=15:5:4:1
Mixed support agent；
(3) 15 parts by weight mycelium, 4 parts by weight adjuvants are mixed with 10 parts by weight wheat straw powder；
(4) 1 parts by weight, 10% imidacloprid wettable powder is added to 38 parts by weight diatomite and 6 weight parts of calcium carbonate
In, it is uniformly mixed；
(5) mixture that step (3), (4) obtain is mixed again, obtains muscardine fine granule.
Embodiment 5
(1) by strain spore inoculating to level liquid culture medium, wherein containing sucrose 3%, peptone 1%, yeast extract powder
0.25%, magnesium sulfate 0.01%, surplus are water, cultivate 48-60 hours, obtain level liquid object；Then it transfers again as strain
To culture medium containing secondary liquid, wherein containing sucrose 2%, soluble starch 1%, brewer's yeast 1%, carrot dry powder 0.4-
0.6%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.05%, surplus are water, cultivate 48-72 hours, obtain secondary liquid culture；
Most of moisture is removed by centrifugation, obtains concentration mycelium；
(2) by weight preparation dialkyl succinylsuccinate sodium sulfonate: hemicellulose: sodium alginate: chitosan=15:5:4:1
Mixed support agent；
(3) 20 parts by weight mycelium, 7 parts by weight adjuvants are mixed with 35 parts by weight Corn core powder；
(4) 1.5 parts by weight, 25% Diacloden wettable powder is added in 46 parts by weight diatomite, is uniformly mixed；
(5) 5 parts by weight carboxymethyl celluloses are uniformly mixed with the mixture of step (3) and step (4), extrusion granulation,
Obtain muscardine granule.
Embodiment 6
(1) by strain spore inoculating to level liquid culture medium, wherein containing sucrose 3%, peptone 1%, yeast extract powder
0.25%, magnesium sulfate 0.01%, surplus are water, cultivate 48-60 hours, obtain level liquid object；Then it transfers again as strain
To culture medium containing secondary liquid, wherein containing sucrose 2%, soluble starch 1%, brewer's yeast 1%, carrot dry powder 0.4-
0.6%, dipotassium hydrogen phosphate 0.1%, magnesium sulfate 0.05%, surplus are water, cultivate 48-72 hours, obtain secondary liquid culture；
Most of moisture is removed by centrifugation, obtains concentration mycelium；
(2) by weight preparation dialkyl succinylsuccinate sodium sulfonate: hemicellulose: sodium alginate: chitosan=15:5:4:1
Mixed support agent；
(3) 25 parts by weight mycelium, 10 parts by weight adjuvants are uniformly mixed with 70 parts by weight wheat straw powder；
(4) 4 parts by weight, 20% Rynaxypyr suspending agent is added to 38 parts by weight attapulgites and 16 parts by weight is white
In carbon black, it is uniformly mixed
(5) mixture of 5 weight account polyethylene alcohol and step (3), step (4) is remixed uniformly, extrusion granulation obtains
Muscardine granule.
The insecticidal toxicity of 7 muscardine preparation of embodiment is tested
Black cutworm (Agrotis ypsilon) larva is acquired from corn field, and insecticidal toxicity survey is carried out in laboratory
It is fixed.Following 4 processing groups: muscardine conidia powder, the muscardine fine granule of above-described embodiment 1,10% pyrrole worm are set
Quinoline wettable powder and control.
Processing step is as follows: 1) it from field fetches soil, is toasted within 2 hours after sieving through 160 DEG C, it is cool to room temperature；2) through surveying
It calculates, the content of conidia powder is 32,000,000,000 spores/g, content is 19.2 hundred million spores/g and imidacloprid containing 0.003g/g in fine granule；3) it weighs
1g conidia powder is soaked and is diluted with 0.1% Tween 80 water of 544mL, is then added in 3199g soil, is puddled uniformly, is obtained
Spore content 1 × 107The processing soil of/g, water content 17:100 (v:w)；4) 20g fine granule is weighed, with 0.1% tween of 649mL
80 water are soaked and are diluted, and are then added in 3820g soil, are puddled uniformly, are obtained spore content 1 × 107/ g, contain 15.6 × 10-6g
The processing soil of imidacloprid/g, water content 17:100 (v:w)；5) 10% imidacloprid wettable powder of 0.468g, 510mL are weighed
It is diluted, is then added in 3000g soil with 0.1% Tween 80 water, puddled uniformly, obtained containing 15.6 × 10-6G imidacloprid/g,
The processing soil of water content 17:100 (v:w)；6) 0.1% Tween 80 water of 510mL is measured, is added in 3000g soil, puddles
Uniformly, control soil is obtained；7) above-mentioned soil is dispensed into respectively in insect box, every box about 50g is respectively put into 1 test worm.Every place
60 (box) is managed, with 20 for 1 group, point 3 groups of repetitions.It is placed in 24-26 DEG C of room temperature to raise 2 weeks, in the 4th, 6,8,10,12,14 day
It checks dead borer population, insecticidal action is evaluated according to the death rate and rate of death, the results are shown in Table 1, Fig. 1.
The toxicity action of 1 muscardine of table and preparation to black cutworm
The result shows that muscardine spore causes the death rate to be 79.3%, LT50It is 8.25 days, and muscardine is micro-
Granula or granule improve the death rate and reach 93.1%, LT50It shorten to 6.46 days, it was demonstrated that bacterial strain has high poison masterpiece to black cutworm
With formula microbial inoculum, which improves, infects rate and the death rate.
The pharmacodynamic test of 8 muscardine preparation of embodiment
It is micro- to the muscardine of above-described embodiment 1-6 with Holotrichia oblita (Holotrichia oblita) 2 instar larvaes
Granula and granule carry out insecticidal toxicity measurement.Following 8 processing are set: muscardine conidia powder, above-described embodiment 1-6
Muscardine fine granule or granule (number be microbial inoculum I, microbial inoculum II, microbial inoculum III, microbial inoculum IV, microbial inoculum V, microbial inoculum VI) and right
According to.
Processing step is as follows: 1) it from field fetches soil, is toasted within 2 hours after sieving through 160 DEG C, it is cool to room temperature；2) through surveying
It calculates, the content of conidia powder is 32,000,000,000 spores/g, 1 content of microbial inoculum is 19.2 hundred million spores/g, 2 content of microbial inoculum is 51.2 hundred million spores/g, microbial inoculum 3
Content is 38.4 hundred million spores/g, 4 content of microbial inoculum is 6.1 hundred million cfu/g, 5 content of microbial inoculum is hundred million spores of 5.3cfu/g, 6 content of microbial inoculum is
Hundred million spores of 4.8cfu/g；3) 1g conidia powder is weighed, is soaked and is diluted with 0.1% Tween 80 water of 544mL, is then added to 3199g soil
In earth, puddles uniformly, obtain spore content 1 × 107The processing soil of/g, water content 17:100 (v:w)；4) parts by weights also known as takes
15.6g microbial inoculum I, 5.9g microbial inoculum II, 7.8g microbial inoculum III, 49.2g microbial inoculum IV, 56.6g microbial inoculum V, 62.5g microbial inoculum VI, it is each to use
0.1% Tween 80 water of 510mL is soaked and is diluted, and is then added in (3000-m) g soil (weight that m is microbial inoculum), is puddled
It is even, obtain content 1 × 107Each microbial inoculum processing soil of/g, water content 17:100 (v:w).6) 0.1% Tween 80 of 510mL is measured
Water is added in 3000g soil, is puddled uniformly, and control soil is obtained；7) above-mentioned soil is dispensed into respectively in insect box, often
Box about 50g is respectively put into 1 test worm.60 (box) of every processing, with 20 for 1 group, point 3 groups of repetitions.24-26 DEG C of room temperature is placed in raise
It supports 2 weeks, in the dead borer population of inspection in the 4th, 6,8,10,12,14 day, insecticidal action is evaluated according to the death rate and rate of death, as a result
It is shown in Table 2.
The toxicity action of 2 muscardine of table and preparation to Holotrichia oblita
The result shows that muscardine spore causes the death rate to be 62.8%, LT50It is 9.35 days, and muscardine is micro-
Granula or granule improve the death rate and reach 71.3~84.1%, LT50It shorten to 6.56 days, it was demonstrated that bacterial strain has Holotrichia oblita
High toxicity action, formula microbial inoculum, which improves, infects rate and the death rate.
The field control effectiveness test of 8 muscardine preparation of embodiment
Chafer larva prevention and control test is carried out in peanut field.Center portion interval is chosen in about 4 mu of peanut ground
The cell for being 21 5m × 5m delimited, 3 cross × 7 extended spreads carry out the following processing: the muscardine particle of above-described embodiment 1
Agent is used by the processing of 100g dosage per acre and the processing of 200g dosage, the muscardine granule of above-described embodiment 4 by 100g per acre
Amount processing and the processing of 200g dosage, 10% imidacloprid wettable powder press the equivalent dosage i.e. per acre 3g in fine granule or granule
With 6g processing, blank control processing.Every processing sets the repetition of 3 cells, is randomly assigned to arrange in cell.Processing method: it is broadcast in peanut
When kind, microbial inoculum or medicament about 1kg fine earth are diluted, spread fertilizer over the fields then dibbling in kind of cave.Planting process routinely unifies pipe
Reason.In harvesting peanut, random 5 points of samplings, 3 continuous plant of every sampling point investigate peanut wormed fruit rate and production in the middle part of every cell
Amount, and check the grub remnants quantity around plant in 20cm square, the function and effect of bacterial strain pest control grub are evaluated, as a result
As shown in table 3.
3 muscardine preparation of table prevents and treats peanut field grub effect
The result shows that field is using after muscardine preparation of the invention, the good fruit number of peanut and weight are significantly improved, worm
Fruit rate and Revision insect recluced rate have raising by a relatively large margin compared with control group.Therefore, Field information muscardine microbial inoculum can be reduced
Grub harm increases peanut yield to reduce wormed fruit rate.
Claims (10)
1. muscardine microbial inoculum, which is characterized in that the muscardine microbial inoculum includes muscardine BRNS50206, mixes
Close adjuvant, mineral carrier and cereal carrier, wherein the deposit number of muscardine BRNS50206 is CGMCC
No.15993。
2. muscardine microbial inoculum according to claim 1, which is characterized in that the muscardine microbial inoculum includes following
Raw material: 5~50 parts of muscardine conidia powder, 4~10 parts of mixed support agent, 20~60 parts of mineral carrier, cereal carry
10~70 parts of body.
3. muscardine microbial inoculum according to claim 1, which is characterized in that the muscardine microbial inoculum includes following
Raw material: 15~25 parts of muscardine mycelium, 4~10 parts of mixed support agent, 20~60 parts of mineral carrier, cereal carry
10~70 parts of body.
4. muscardine microbial inoculum according to claim 1, which is characterized in that mixed support agent is dialkyl succinylsuccinate sulfonic acid
Sodium, hemicellulose, sodium alginate, chitosan are 15:5:4:1 mixing according to mass ratio.
5. muscardine microbial inoculum according to claim 1, which is characterized in that the mineral carrier is attapulgite, cunning
One of mountain flour, diatomite, white carbon black, calcium carbonate, clay are a variety of.
6. muscardine microbial inoculum according to claim 1, which is characterized in that the cereal carrier be peanut pod shell powder,
One of maize cob meal, corn stalk powder, wheat straw powder, rice straw powder, sugar beet powder, bagasse powder are a variety of.
7. muscardine microbial inoculum according to claim 1, which is characterized in that the muscardine microbial inoculum further includes gluing
0.1~6 part of mixture, described adhesive is polyvinyl alcohol or carboxymethyl cellulose.
8. muscardine microbial inoculum described in any one according to claim 1~7, which is characterized in that the muscardine
Microbial inoculum further includes 1~4 part of synergist.
9. muscardine microbial inoculum according to claim 8, which is characterized in that the synergist is that 10% imidacloprid is wettable
Property pulvis, 25% Diacloden wettable powder or 20% Rynaxypyr suspending agent.
10. the application of muscardine microbial inoculum described in claim 1.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113481109A (en) * | 2021-07-30 | 2021-10-08 | 沧州市农林科学院 | Beauveria bassiana and application thereof in preventing and treating scarab beetles |
| CN113880660A (en) * | 2021-09-17 | 2022-01-04 | 湖北问心农业科技有限公司 | Biological organic fertilizer for killing underground insect pests by using biological agent and preparation method thereof |
| CN115231974A (en) * | 2022-07-22 | 2022-10-25 | 黑龙江希碧莱生物工程技术有限公司 | Crop bacterial fertilizer for preventing and treating soil insects and grubs and preparation method thereof |
-
2019
- 2019-02-10 CN CN201910109413.6A patent/CN109757521B/en active Active
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113481109A (en) * | 2021-07-30 | 2021-10-08 | 沧州市农林科学院 | Beauveria bassiana and application thereof in preventing and treating scarab beetles |
| CN113880660A (en) * | 2021-09-17 | 2022-01-04 | 湖北问心农业科技有限公司 | Biological organic fertilizer for killing underground insect pests by using biological agent and preparation method thereof |
| CN115231974A (en) * | 2022-07-22 | 2022-10-25 | 黑龙江希碧莱生物工程技术有限公司 | Crop bacterial fertilizer for preventing and treating soil insects and grubs and preparation method thereof |
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