CN109769861A - Metarhizium anisopliae IPPMHBC-009 composite bacteria agent and its application - Google Patents
Metarhizium anisopliae IPPMHBC-009 composite bacteria agent and its application Download PDFInfo
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- CN109769861A CN109769861A CN201910187082.8A CN201910187082A CN109769861A CN 109769861 A CN109769861 A CN 109769861A CN 201910187082 A CN201910187082 A CN 201910187082A CN 109769861 A CN109769861 A CN 109769861A
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Abstract
The invention belongs to field of biotechnology, and in particular to a kind of Metarhizium anisopliae IPPMHBC-009 composite bacteria agent and its application.Green muscardine fungus (Metarhizium anisopliae) IPPMHBC-009, deposit number are CGMCC No.16000, have good LD50 time and bombys batryticatus rate for Oedaleus asiaticus B, Dasyhippus barbipes.Green muscardine fungus IPPMHBC-009 and Rynaxypyr are administered in combination the present invention, can significantly improve the quick-acting and killing rate of desinsection.
Description
Technical field
The invention belongs to field of biotechnology, and in particular to a kind of Metarhizium anisopliae IPPMHBC-009 composite bacteria agent and
Using.
Background technique
Locust is the disastrous insect of Orthoptera locust section, and the plague of locusts frequently occurs in China, causes to China's agricultural huge
Injury.Currently, China control grassland grasshopper harm means mainly use chemical prevention method, have quickly, efficiently, make
With it is flexible the advantages that, still, as people increasingly pay attention to the sustainable development of environmental economy, biological locust elimination method is developed
Come and applies in real work.Biology goes out locust method, especially green muscardine fungus, muscardine etc., has certain specificity, to people
It raises harmless, while also having many advantages, such as that free from environmental pollution, noresidue, pest will not develop drug resistance.
Although disinsection fungal, which prevents and treats locust, has certain control efficiency, fungi is knocked down, killing off the insect pests acts on slowly,
It needs just be worked after a period of time, is suitble to general generation time control and application, and in locust outbreak year part, or part
When regional serious generation, disinsection fungal cannot quickly reduce insect density in the short time.Therefore, how to utilize disinsection fungal short
The quantity of control locust is agriculturally urgent problem to be solved in time.
Bacterial strain and agriculture chemical compounding are that raising locust control effect explores approach, but bacterial strain and pesticide at present
Compounding effect is unstable, and compounding is improper to fail to improve instead even reduction locust control effect.
Summary of the invention
In order to obtain effective bacterial strain and agriculture chemical compounding formula, the present invention provides a kind of Metarhizium anisopliae IPPMHBC-
009 composite bacteria agent and its application.
The object of the present invention is to provide the compound bacterias comprising Metarhizium anisopliae IPPMHBC-009 and Rynaxypyr
Agent.
A further object of the present invention is to provide the applications of Metarhizium anisopliae IPPMHBC-009 composite bacteria agent.
A further object of the present invention is to provide use Metarhizium anisopliae IPPMHBC-009 composite bacteria agent to carry out anti-locust elimination
The method of worm.
Metarhizium anisopliae (Metarhizium anisopliae) IPPMHBC- of specific embodiment according to the present invention
The deposit number CGMCC No.16000 of 009 bacterial strain, the bacterial strain were stored in China General Microbiological bacterium on 08 02nd, 2018
Kind collection, collection address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica.
The composite bacteria agent of the efficient killing locust of specific embodiment according to the present invention, it includes above-mentioned Metarhizium anisopliaes
IPPMHBC-009 and Rynaxypyr.
The composite bacteria agent of the efficient killing locust of specific embodiment according to the present invention, Rynaxypyr and chafer are green
The mass ratio of stiff bacterium IPPMHBC-009 is 1:1000~2000.
The microbial inoculum of the efficient killing locust of specific embodiment according to the present invention, the concentration of Rynaxypyr are 0.004
The mass concentration of~0.1ppm, Metarhizium anisopliae IPPMHBC-009 are 0.0064~0.16%.When with bacteriostat, chlorine worm benzene
Formamide is diluted to normal concentration with acetone, and Metarhizium anisopliae IPPMHBC-009 conidia powder is mixed with wheat bran according to mass ratio
It is configured to normal concentration.
Metarhizium anisopliae IPPMHBC-009 of the invention is 3.87d, bombys batryticatus rate to the median lethal time of Dasyhippus barbipes
It is 91.67%, the median lethal time to Oedaleus asiaticus B is 3.55d, and bombys batryticatus rate is 93.33%, therefore, green muscardine fungus of the invention
IPPMHBC-009 is with good application prospect.
Metarhizium anisopliae IPPMHBC-009 and Rynaxypyr combination can be mentioned significantly and be killed locust by the present invention
It is lethal slow not only to solve the problems, such as fungus insecticide for quick-acting and killing rate, and largely reduce pollution, alleviate
Pest develops drug resistance problem to chemical pesticide.
Detailed description of the invention
Fig. 1 shows 9 kinds of green muscardine fungus to the correction cumulative mortality curve of Dasyhippus barbipes 15d；
Fig. 2 shows 9 kinds of green muscardine fungus to the correction cumulative mortality curve of Oedaleus asiaticus B 15d；
Fig. 3 shows the cumulative mortality of Oedaleus asiaticus B after processing, A: only application Rynaxypyr；B: it only applies green
Stiff bacterium IPPMHBC-009；C: mixing Rynaxypyr and green muscardine fungus IPPMHBC-009.
The deposit number CGMCC of Metarhizium anisopliae (Metarhizium anisopliae) IPPMHBC-009 bacterial strain
No.16000 was stored in China General Microbiological Culture Collection Center, collection address: Beijing on 08 02nd, 2018
The Institute of Microorganism, Academia Sinica, institute of Chaoyang District North Star West Road 1.
Specific embodiment
The source of test worm of the present invention:
1. Dasyhippus barbipes (Dasyhippus barbipes Fischer-Waldheim): 2~3 age nymphs of a locust, method of beating
It is collected in In Xilingol League In Inner Mongolia white clouds coulomb locust area.
2. Oedaleus asiaticus B (Oedaleus decorus asiaticus B.Bienko): 2~3 age nymphs of a locust, the method for beating are adopted
Combine in the In Xilingol League In Inner Mongolia west northern 7 kilometers of areas Chu Huang of black flag Bayan Gol.
The locust raising of acquisition feeds suitable fresh sheep's hay in the big frame in experiment station dedicated for supporting locust daily
Blade.For guarantee test effect, test the previous day Nature enemy.
Bioassay of the 1 Metarhizium anisopliae IPPMHBC-009 of embodiment to Dasyhippus barbipes nymph of a locust's pathogenicity
Metarhizium anisopliae IPPMHBC-009 is isolated from soil, and deposit number is CGMCC No.16000.
9 kinds of Metarhizium Strains are measured to the virulence effect of locust using stomach toxicity method.
0.2g muscardine spore powder is weighed first to be added in 2.0g wheat bran, adds 0.1ml edible oil, is sufficiently stirred
It is even.Then it weighs the green muscardine fungus wheat bran bait formulation that 0.2g is prepared to be put into culture dish, is put into togerther cover glass plate together with culture dish
Worm basket, number are supported, then is put into 20 3 age Dasyhippus barbipes nymphs of a locust of the same size in feeding worm basket.Blank control group wheat bran bait formulation
In muscardine spore powder is not added.The present invention selects 9 plants of muscardine spore powder and a control CK, every kind of conidia powder to be arranged 3 times
It repeats, i.e., totally 33 processing, in being raised in insectary.Bait formulation was taken out in second day, feeds suitable fresh sheep's hay daily later,
And the death condition of every basket of nymph of a locust is investigated, it makes a record respectively.
The dead nymph of a locust is sorted out and is respectively put into the culture dish for being lined with filter paper of reference numeral, certain humidity is kept, makes
Grow bombys batryticatus.Every kind of bacterium is investigated after the nymph of a locust to be tested is all dead treated the bombys batryticatus rate of the nymph of a locust, 15 days after combination processing
Interior daily cumulative mortality and median lethal time (LT50) pick out to the preferable strain of locust lethal effect.
9 plants of Metarhizium Strains are shown in figure l, table 1 to the bioassay results of 3 age Dasyhippus barbipes nymph of a locust pathogenicities.
Comparison of the 19 plants of green muscardine fungus of table to Dasyhippus barbipes pathogenicity
Analyzed by table 1 it is found that above-mentioned 9 kinds of strains testeds have to the nymph of a locust it is pathogenic, median lethal time 3.87d~
Between 8.83d, bombys batryticatus rate is between 43.33%~91.67%.The wherein median lethal time of Metarhizium Strains IPPMHBC-009
For 3.87d, bombys batryticatus rate is 91.67%, and 803,200614, the bacterial strains median lethal time such as 2010-10 is slightly long, bombys batryticatus rate is lower.
Therefore, compared with the bacterial strains such as 803,200614, there are significant differences by Metarhizium Strains IPPMHBC-009, to Dasyhippus barbipes
There is higher pathogenicity.
Bioassay of the 2. Metarhizium anisopliae IPPMHBC-009 of embodiment to Oedaleus asiaticus B pathogenicity
9 plants of bacterial strains are measured to the power of curing the disease of Oedaleus asiaticus B, method of the measuring method with embodiment 1.
9 plants of Metarhizium Strains are shown in Fig. 2, table 2 to the bioassay results of Oedaleus asiaticus B pathogenicity.
Comparison of the 29 plants of green muscardine fungus of table to Oedaleus asiaticus B pathogenicity
Analyzed by table 2 it is found that strains tested is to the nymph of a locust have it is pathogenic, median lethal time between 3.55d~10.67d,
Bombys batryticatus rate is between 38.33%~93.33%.Wherein the median lethal time of Metarhizium Strains IPPMHBC-009 is 3.55d, stiff
Worm rate is 93.33%.Compared with bacterial strain 200614 etc., Metarhizium Strains IPPMHBC-009 has significant difference, shows it to Asia
Trolley locust has higher pathogenicity in continent.
Embodiment 3 measures Metarhizium anisopliae IPPMHBC-009 and is compounded with chemical pesticide to Oedaleus asiaticus B pathogenicity
Test sets 3 processing, and every processing includes 3 groups of the 3 age nymph of a locust, every group of 3 repetitions.1st processing low quality concentration
95.5% Rynaxypyr (0.004,0.02,0.1ppm), the 2nd processing with green muscardine fungus IPPMHBC-009 handle
(0.0064%, 0.032%, 0.16%), the 3rd processing 95.5% Rynaxypyr of low concentration and green muscardine fungus IPPMHBC-
009 mixed liquor (0.004ppm+0.0064%, 0.02ppm+0.032%, 0.1ppm+0.16%).
Processing method: the first processing group dilutes 95.5% Rynaxypyr with acetone, obtain 0.004,0.02,
0.1ppm；Second processing group dilutes green muscardine fungus IPPMHBC-009 conidia powder with wheat bran, obtain 0.0064%, 0.032%,
0.16% 3 concentration；Third processing group compounds the first processing processing group with second processing group, obtains.
Test uses stomach toxicity method.It takes the bait formulation 0.2g prepared to be put into culture dish, is respectively put into together with culture dish each
It supports in worm basket, number.Each processing puts 15 for trying the 3 age nymphs of a locust, is placed in a feeding worm basket, feeds fresh sheep's hay daily.
Every processing is repeated 3 times.Daily observation 1 time, and observe and record.
Oedaleus asiaticus B is tired when Rynaxypyr, green muscardine fungus IPPMHBC-009 and the mixing application of bacterium medicine is administered alone
The death rate is counted as shown in Fig. 3, table 3.
Comparison of the 3 green muscardine fungus microbial inoculum of table to Oedaleus asiaticus B pathogenicity
As a result as shown in table 3, Fig. 3, after green muscardine fungus IPPMHBC-009 is used in mixed way with Rynaxypyr, in quick-acting
It all improves a lot in terms of killing rate, not only reduces the spore germination rate of green muscardine fungus to a certain extent, and substantially reduce
The immunity of pest is allowed to more easily by green muscardine fungus parasitism.Therefore, green muscardine fungus IPPMHBC-009 and Rynaxypyr are mixed
The advantages of application can be in conjunction with both sides are closed, on the one hand can solve the lethal slow problem of fungus insecticide, it on the other hand can be with
Pollution is largely reduced, alleviates pest and develops drug resistance problem to chemical pesticide, to play synergistic purpose.
After green muscardine fungus IPPMHBC-009 of the invention is compounded with other chemical pesticides in addition to Rynaxypyr, locust
Worm control efficiency is not simultaneously significant, or even the case where not rising anti-drop occurs.
Claims (4)
1. Metarhizium anisopliae IPPMHBC-009 composite bacteria agent, which is characterized in that the composite bacteria agent includes Rynaxypyr
With Metarhizium anisopliae IPPMHBC-009, the deposit number of the Metarhizium anisopliae IPPMHBC-009 is CGMCC
No.16000。
2. Metarhizium anisopliae IPPMHBC-009 composite bacteria agent according to claim 1, which is characterized in that described compound
The mass ratio of Rynaxypyr and Metarhizium anisopliae IPPMHBC-009 are 1:1000~2000 in microbial inoculum.
3. application of the Metarhizium anisopliae IPPMHBC-009 composite bacteria agent described in claim 1 in terms of preventing and treating locust.
4. a kind of method for preventing and treating locust, which is characterized in that the method includes to use the green deadlock of chafer described in claim 1
Bacterium IPPMHBC-009 composite bacteria agent prevents and treats the step of locust.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112063602A (en) * | 2020-09-03 | 2020-12-11 | 宁夏中微泰克生物技术有限责任公司 | Asian locusta migratoria small G protein Ras and coding gene and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104865350A (en) * | 2015-05-15 | 2015-08-26 | 中国农业科学院植物保护研究所 | Insecticidal toxicity evaluation method of green muscadine fungus and chlorantraniliprole combined utilization |
| CN109337823A (en) * | 2018-10-11 | 2019-02-15 | 中国农业科学院植物保护研究所 | Metarhizium anisopliae IPPMHBC-009 and its application |
-
2019
- 2019-03-13 CN CN201910187082.8A patent/CN109769861A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104865350A (en) * | 2015-05-15 | 2015-08-26 | 中国农业科学院植物保护研究所 | Insecticidal toxicity evaluation method of green muscadine fungus and chlorantraniliprole combined utilization |
| CN109337823A (en) * | 2018-10-11 | 2019-02-15 | 中国农业科学院植物保护研究所 | Metarhizium anisopliae IPPMHBC-009 and its application |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112063602A (en) * | 2020-09-03 | 2020-12-11 | 宁夏中微泰克生物技术有限责任公司 | Asian locusta migratoria small G protein Ras and coding gene and application thereof |
| CN112063602B (en) * | 2020-09-03 | 2022-10-21 | 宁夏中微泰克生物技术有限责任公司 | Asian locusta migratoria small G protein Ras and coding gene and application thereof |
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Application publication date: 20190521 |