CN104611240A - Application of Metarhizium anisopliae strain in preparation of insecticide for vegetable pest insects - Google Patents
Application of Metarhizium anisopliae strain in preparation of insecticide for vegetable pest insects Download PDFInfo
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Abstract
The invention relates to an application of a Metarhizium anisopliae CQMa421 strain in preparation of insecticide for preventing and controlling Striped flea beetle or plutella xylostella. The strain is capable of infecting Striped flea beetles and plutella xylostella with fatality rates being 60.9% and 70.8% respectively. The Metarhizium anisopliae CQMa421 strain with enhanced toxicity, which is prepared according to a method disclosed by the invention, has a fatality rate reaching 100% and achieves an outstanding prevention and control effect. The insecticide for preventing and controlling Striped flea beetles or plutella xylostella comprises dry conidial powder of Metarhizium anisopliae CQMA421 (44-60%), kieselguhr (30-40%), nano-aluminum trihydrate (0.5-1%), sodium carboxymethylcellulose (3-5%), tween-80 (3-5%) and OP-10 (3-5%). All the components in the Metarhizium anisopliae insecticide are matched effectively to take a synergistic effect. After being mixed with water, the Metarhizium anisopliae insecticide is applied in a spraying way to achieve a field prevention effect of 80% or above for rice planthopper. The Metarhizium anisopliae insecticide has excellent wetting dispersion characteristic, wherein the wetting time is less than 60s; and in addition, the bioactivity during long-term storage can be retained and the storage time is as long as 12 months.
Description
Technical field
The present invention relates to a kind of new opplication of Metarhizium anisopliae bacterial strain, be specifically related to one and kill vegetables pest Phyllotreta striolata, small cabbage moth Metarhizium anisopliae preparation and application thereof, belong to field of biological pesticide.
Background technology
Phyllotreta striolata, belongs to Coleoptera, Chrysomelidae insect, and being called for short flea beetle, is leaf vegetables important pests.Small cabbage moth, is commonly called as little blue or green worm, leaf vegetables of also mainly causing harm.Both insect often occurs at vegetable field simultaneously or alternately, brings massive losses every year to Vegetable produce.At present, mainly use chemical pesticide and control Phyllotreta striolata and small cabbage moth, this has a strong impact on HUMAN HEALTH, destroys ecotope.Therefore the microbial preparation of employing safety substitutes the important development direction that chemical pesticide becomes Phyllotreta striolata and small cabbage moth control.
Insect pathogenic fungus is most important insecticidal microorganism monoid, and fungus insecticide, as important alternative agricultural chemicals, is subject to extensive concern in recent years.Green muscardine fungus (
metarhizium) be that can infect the whole world about has 200 various insects the earliest for one of the insect pathogenic fungus of pest control, utilize it to carry out the Researching and practicing of pest control more century-old.Be separated to the disinsection fungal bacterial strain that some can infect Phyllotreta striolata or small cabbage moth at present, the preparation of part bacterial strain is registered control small cabbage moth, but could not be used widely, and mainly because bacterial strain insecticidal range is narrow, can not prevent and treat other insects of vegetables simultaneously.Metarhizium anisopliae CQMa421 be the strain production traits that a strain separates from the Cnaphalocrocis medinali(rice leaf roller) bombys batryticatus of natural infection good, to Cnaphalocrocis medinali(rice leaf roller), there is high virulence disinsection fungal, open in Chinese patent CN102212483, this bacterial strain on 02 25th, 2011 in China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC) preservation, deposit number: CGMCC NO.4609.This bacterial strain and preparation thereof have no report to the virulence of Phyllotreta striolata and small cabbage moth and prevention effect.
Wettable powder is the most basic a kind of pesticide formulation, can be made into suspension liquid, sprays.Disinsection fungal wettable powder adds a certain amount of tensio-active agent and stopping composition by fungal spore powder, makes by sieving.Because fungal spore activity is by the impact of wetting agent, often reduce the consumption of tensio-active agent in preparation, cause moistening dispersion effect poor, product is still shorter for storage period simultaneously, rarely exceeds 12 months.Because the problem of moistening dispersing characteristic, package stability and field efficacy affect greatly by uv irradiating, though disinsection fungal wettable powder has registration, never widely apply.Therefore, screen wide spectrum bacterial strain, tame supper toxic strain and develop the good fungus insecticide wettable powder of moistening dispersing characteristic, extend its package stability, thus promote the application of fungus insecticide in control Phyllotreta striolata and small cabbage moth, be conducive to the consumption reducing chemical pesticide in Rice Production.
Summary of the invention
The first object of the present invention is to provide a kind of Metarhizium anisopliae bacterial strain killing the application in Phyllotreta striolata or small cabbage moth.
The present invention second object is to provide a kind of Metarhizium anisopliae preparation, and the resistance to uv irradiating of said preparation, long preservative period, preventive effect are good.
The present invention the 3rd object is that providing a kind of strengthens Metarhizium anisopliae virulence, and does not affect the method that it produces spore, heat-resistant quality.
The present invention seeks to be achieved through the following technical solutions:
The application of Metarhizium anisopliae CQMa421 bacterial strain in the medicament preparing control Phyllotreta striolata or small cabbage moth, this bacterial strain can infect Phyllotreta striolata and small cabbage moth, and lethality rate can reach 60.9% and 70.8% respectively.
Different Metarhizium Strains has specific host, as open report: Metarhizium anisopliae FI-1045 bacterial strain can infect sugarcane grub (Anoplophora glabripennis), Metarhizium anisopliae A ESALQ818 bacterial strain can infect Aedes Aegypti (Aedes aegypti), Metarhizium anisopliae EAMb 09/01-Su bacterial strain can infect Mediterranean Sea fruit fly, Metarhizium anisopliae PDRL526 bacterial strain can infect radish aphid (Lipaphis erysimi), Metarhizium anisopliae M2 bacterial strain can infect Aleurodicus dispersus (Aleurodicus dispersus Russell), Metarhizium anisopliae ICIPE-30 bacterial strain can infect anopheles costalis (anopheles gambiae), Metarhizium anisopliae ESC 1 bacterial strain can infect termite, Metarhizium anisopliae ATCC62176 can infect beet root maggot (sugarbeet root maggot), Metarhizium anisopliae M105 bacterial strain can infect locust, Metarhizium anisopliae wide spectrum bacterial strain F-52 can infect grub (Anoplophora glabripennis), walnut weevil, Xylosandrus germanus (Xylosandrus germanus), Metarhizium anisopliae wide spectrum bacterial strain 549 can infect maduca sexta (Manduca sexta), Aedes Aegypti (Aedes aegypti), Luo Baici green muscardine fungus wide spectrum bacterial strain ARSEF 23 can infect locust, lepidopterous larvae, beetle etc.
Preferably, the medicament of above-mentioned control Phyllotreta striolata or small cabbage moth is made up of, all with weight percent green muscardine fungus CQMa421 xerospore powder 44-60%, diatomite 30-40%, nano-aluminum hydroxide 0.5-1%, Xylo-Mucine 3-5%, tween-80 3-5% and OP-10 3-5%.
A kind of Metarhizium anisopliae preparation, it is characterized in that: it is made up of, all with weight percent diatomite 30-40%, nano-aluminum hydroxide 0.5-1%, Xylo-Mucine 3-5%, tween-80 3-5%, OP-10 3-5% and green muscardine fungus CQMa421 xerospore powder 44-60%.
Above-mentioned each component is the clear and definite component in this area, tween-80 is polyoxyethylene sorbitan monooleate, is called for short Tween-80, OP-10 is alkylphenol polyoxyethylene, green muscardine fungus CQMa421 is open in Chinese patent CN102212483, this bacterial strain on 02 25th, 2011 at China Committee for Culture Collection of Microorganisms's common micro-organisms center (CGMCC) preservation, deposit number: CGMCC NO.4609; Nano-aluminum hydroxide is particle diameter is nano level aluminium hydroxide.
The tween-80 of above-mentioned tween-80 and the preferred purity >95% of OP-10 and OP-10; Above-mentioned Xylo-Mucine (CMC) preferably purity is greater than 95%; Above-mentioned diatomite is preferably white to light brown loose powder, and purity is greater than 85%, and water content is less than 1%; Above-mentioned nano-aluminum hydroxide is preferably particle diameter and is less than 30nm.Above-mentioned green muscardine fungus CQMa421 xerospore powder is pure spore powder, spore content >=15,000,000,000 spores/gram, water content≤5%; Above all in mass percentage.
In above-mentioned Metarhizium anisopliae preparation, nano-aluminum hydroxide is attached to fungal spore hydrophobic surface, has effectively intercepted the detrimentally affect of preparation middle and high concentration (3-5wt%) tween-80 and OP-10, effectively ensure that the biological activity of spore.
Further preferably, Metarhizium anisopliae preparation of the present invention, it is characterized in that: it is made up of, all with weight percent diatomite 35-40%, nano-aluminum hydroxide 0.5-0.8%, Xylo-Mucine 3-5%, tween-80 3-5%, OP-10 3-5% and green muscardine fungus CQMa421 xerospore powder 45-55%.
Above-mentioned green muscardine fungus CQMa421 xerospore powder preferably adopts strong virus force Metarhizium anisopliae CQMa421 spore powder, described strong virus force Metarhizium anisopliae CQMa421 spore powder is obtained by following methods: 28 DEG C, in humidity 50-70% environment, Metarhizium anisopliae CQMa421 is inoculated in Phyllotreta striolata imago breeding, select the Phyllotreta striolata of susceptible death at first, put into 28 DEG C of incubator moisturizings and cultivate 3-5 days, make its insect surfaces grow Fresh spores; Collect Fresh spores, inoculation 3-4 diamondback moth larvae in age, the Fresh spores that the same acquisition insect surfaces grows.Circulation inoculation Phyllotreta striolata described above and small cabbage moth obtain the Metarhizium anisopliae CQMa421 Fresh spores after taming for 16 times; Aseptically, ruling containing on the 1/4SDA culture medium flat plate of 100 mcg/ml penbritins with Fresh spores after the domestication of transfering loop picking, the culture dish of having rule upset is being positioned in 28 DEG C of thermostat containers and cultivates; The typical case that grows is selected and without the bacterium colony of miscellaneous bacteria after 3-4 days, at colony edge with moving substratum one fritter of acicula picking with mycelia, proceed to test tube slant substratum central authorities, 28 DEG C of Metarhizium anisopliae CQMa421 mothers obtaining strong virus force for constant temperature culture 10-15 days plant, CQMa421 bacterial strain mother being planted is seeded on 1/4SDA flat board, cultivate 15 days, collect ripe conidium and obtain green muscardine fungus CQMa421 xerospore powder for 28 DEG C.
The preparation method of above-mentioned Metarhizium anisopliae preparation, first green muscardine fungus CQMa421 xerospore powder and nano-aluminum hydroxide are mixed in mixing machine, add diatomite, Xylo-Mucine, tween-80 and OP-10 successively again, stir while adding, until each component mixes obtained completely.
A kind of method strengthening Metarhizium anisopliae virulence, it is characterized in that: 28 DEG C, in humidity 50-70% environment, Metarhizium anisopliae CQMa421 is inoculated in Phyllotreta striolata imago breeding, select the Phyllotreta striolata of susceptible death at first, put into 28 DEG C of incubator moisturizings and cultivate 3-5 days, make its insect surfaces grow Fresh spores; Collect Fresh spores, inoculation 3-4 diamondback moth larvae in age, the Fresh spores that the same acquisition insect surfaces grows.Circulation inoculation Phyllotreta striolata described above and small cabbage moth obtain the Metarhizium anisopliae CQMa421 Fresh spores after taming for 16 times; Aseptically, ruling containing on the 1/4SDA culture medium flat plate of 100 mcg/ml penbritins with Fresh spores after the domestication of transfering loop picking, the culture dish of having rule upset is being positioned in 28 DEG C of thermostat containers and cultivates; The typical case that grows is selected and without the bacterium colony of miscellaneous bacteria after 3-4 days, at colony edge with moving substratum one fritter of acicula picking with mycelia, proceed to test tube slant substratum central authorities, 28 DEG C of Metarhizium anisopliae CQMa421 mothers obtaining strong virus force for constant temperature culture 10-15 days plant.
Strong virus force Metarhizium anisopliae CQMa421 bacterial strain after adopting the present invention to strengthen the method domestication of virulence, can improve 39.1% and 29.2% respectively to Phyllotreta striolata and small cabbage moth lethality rate, reach 100%, effect is very outstanding.
The present invention has following beneficial effect:
Metarhizium anisopliae CQMa421 bacterial strain of the present invention can have and infects Phyllotreta striolata, small cabbage moth, lethality rate can reach 39.1% and 29.2% respectively, the Metarhizium anisopliae CQMa421 bacterial strain that the method adopting the present invention to strengthen virulence obtains, its lethality rate all can reach 100%, and effect is very outstanding.In Metarhizium anisopliae preparation of the present invention each composition effective cooperation, serve synergistic function; Metarhizium anisopliae preparation of the present invention is adopted to be watered spraying; all more than 80% is reached to the field efficacy of Phyllotreta striolata and small cabbage moth; said preparation has good uvioresistant and wetting and dispersing characteristic, wetting time < 60s; and the biological activity maintained in standing storage; period of storage can reach 12 months, is very suitable for large-scale production and spread use.
Embodiment
Below by embodiment, the present invention is specifically described; what be necessary to herein means out is that following examples are only used to further illustrate the present invention; limiting the scope of the invention can not be interpreted as; without departing from the spirit and substance of the case in the present invention; the amendment do the inventive method, step or condition or replacement, all belong to scope of the present invention.
If do not specialize, the conventional means that technique means used in embodiment is well known to those skilled in the art.
embodiment 1cQMa421 bacterial strain and the virulence to Phyllotreta striolata and small cabbage moth after inoculation Phyllotreta striolata and the domestication of small cabbage moth virulence
By CQMa421 inoculation on 1/4SDA flat board, cultivate 15 days, collect ripe conidium, with 0.05%(W/V for 28 DEG C) tween-80 dispersion, being mixed with final concentration is 1 × 10
7the suspension of individual spore/ml.The Phyllotreta striolata adult of indoor feeding being proceeded to length has in the 500ml triangular flask of cabbage leaves, and every bottle of 30-50 only, adopts spray tower (POTTER, BURKARD company) spray inoculation, inoculates 5 bottles, every bottle graft kind 0.5 ml.Room temperature 28 DEG C after inoculation, in the raw side room of humidity 50-70%; Select the Phyllotreta striolata of susceptible death at first, put into 28 DEG C of incubator moisturizings and cultivate 3-5 days, make its insect surfaces grow Fresh spores.It is 1 × 10 that collection spore is mixed with final concentration again
7the suspension of individual spore/ml, adopts topical application inoculation 3-4 diamondback moth larvae in age, and every examination worm micro-sampling device drop medicament 2 μ l, inoculates 100, room temperature 28 DEG C after inoculation, raise under the condition of humidity 50-70%; Select the small cabbage moth bombys batryticatus of susceptible death at first, put into 28 DEG C of incubator moisturizings and cultivate 3-5 days, make its insect surfaces grow Fresh spores.Circulation inoculation Phyllotreta striolata described above and small cabbage moth are tamed for 15-17 time.Aseptically, ruling containing on the 1/4SDA culture medium flat plate of 100 mcg/ml penbritins with Fresh spores after the domestication of transfering loop picking; The culture dish of having rule upset is positioned in 28 DEG C of thermostat containers and cultivates; Select the typical case that grows after 3-4 days and without the bacterium colony of miscellaneous bacteria, at colony edge with moving substratum one fritter of acicula picking with mycelia, proceed to test tube slant substratum central authorities, 28 DEG C of constant temperature culture 10-15 days.Bacterial strain after domestication cultivates conidium as stated above, inoculates Phyllotreta striolata and small cabbage moth respectively, to tame front bacterial strain for contrast, repeats 3 times.Record dead borer population, calculate cumulative mortality, carry out statistical study through SPSS software.
Sporulating character: get the spore suspension 100 μ l preparing CQMa421 and be seeded on 1/4SDA flat board, cultivate 15 days under the culture condition of 28 DEG C of temperature, beat at random with the punch tool that diameter is 1cm and get 5 bacterium blocks, drop into and the 0.05%(W/V of 50ml be housed) disperse in the triangular flask of tween-80, blood counting chamber is utilized to measure spore concentration, the sporulation quantity (spore count/cm on Units of Account area
2), to tame front bacterial strain for contrast, repeat 3 times.Statistical study is carried out with SPSS software.
Heat-resistant quality: the spore suspension preparing CQMa421 is processed 5 hours respectively at 50 DEG C, respectively gets 100 μ l and be seeded on 1/4SDA flat board, cultivate at 28 DEG C of temperature.If nonheat-treated conidiospore suspension is contrast.Three repetitions are established in each process.After 24 hours, sediments microscope inspection, several 200-300 spores, the spore count that record is sprouted, calculates germination rate.To tame front bacterial strain for contrast, repeat 3 times.Statistical study is carried out with SPSS software.
Virulence, sporulating character and the heat-resistant quality of table 1 CQMa421 bacterial strain after the domestication of inoculation rice leafhopper
Note: in the level of p=0.01, same letter represents that difference is not remarkable, not same letter significant difference.
From can table 1 to find out, CQMa421 can effectively infect Phyllotreta striolata and small cabbage moth, inoculates latter 8 days, and CQMa421 treatment group mortality ratio is respectively 60.9% and 70.8%; Significantly strengthen the insecticidal activity of Phyllotreta striolata and small cabbage moth after the domestication of CQMa421 bacterial strain after taming 16 times, lethality rate all reaches 100%, comparatively improves 39.1% and 29.2% respectively before domestication; But to there was no significant difference before the virulence of Cnaphalocrocis medinali(rice leaf roller) and the sporulation quantity on 1/4SDA flat board, thermotolerance and domestication after the domestication of CQMa421 bacterial strain, illustrate, after Phyllotreta striolata and small cabbage moth circulation domestication 16 times, spore and thermotolerance are produced without impact to bacterial strain CQMa421.
embodiment 2nano-aluminum hydroxide is to green muscardine fungus provide protection
After pure for Metarhizium anisopliae CQMa421 spore powder first being mixed with nano-aluminum hydroxide by table 2, then add tween-80, the OP-10 mixing of 40% diatomite, 5%CMC and different content.
Ultraviolet protection effect: it is 1 × 10 that the water-dispersion of preparation sample is made into concentration
7after the spore suspension of spore/mL, get 100 μ l spore suspensions and be applied on 1/4 SDA flat board, with glass push away even after be placed in 290-310nm ultraviolet lamp (978mW/m
2) under vertical irradiation 5 hours, then be placed in 28 DEG C of thermostat containers and cultivate 24h hour, measure germination rate (method is the same), repeat 3 times.With SPSS software, Treatment Analysis is carried out to data.
Store experiment: preserve 2 months at 28 DEG C of lower seals by after preparation sample, being made into concentration with water-dispersion is 1 × 10
7after the spore suspension of spore/mL, measure germination rate (method is the same), repeat 3 times.With SPSS software, Treatment Analysis is carried out to data.
Table 2 nano-aluminum hydroxide is to green muscardine fungus provide protection
The results are shown in Table 2, the nano-aluminum hydroxide of 0.5-1% has obvious provide protection to Metarhizium anisopliae CQMa421 spore, when tween-80 and OP-10 content are 3 ~ 5%, the spore of storage preservation after 2 months is active more than 90%, is significantly higher than the process of non-plus nano aluminium hydroxide; Also spore uvioresistant ability is significantly increased, for field efficacy stability lays the foundation.
embodiment 3the preparation of Metarhizium anisopliae CQMa421 wettable powder and wettability, storage period and preventive effect.
After first being mixed with nano-aluminum hydroxide by pure for Metarhizium anisopliae CQMa421 cryptogam by table 2, then add different content diatomite, CMC, tween-80 and OP-10.
The wettability of Ricinate measures: get standard water 100mL and inject 250 mL beakers.This beaker is placed in the water bath with thermostatic control of 25 DEG C ± 1 DEG C, makes its liquid level concordant with the horizontal plane of water-bath.When hard water to 25 DEG C ± 1 DEG C, accurately take 5.0g testing sample and be placed on watch-glass, whole sample is poured over the liquid level of this beaker equably from the position flushed with beaker mouth is disposable, but not disturbance liquid level excessively.Clock with stopwatch immediately when adding sample, till sample is all wetting (the fine powder film stayed on liquid level can be ignored).Write down wetting time (being accurate to second).Repetition like this 5 times, gets its mean value, as the wetting time of this sample.
Package stability: preparation is preserved 12 months at 28 DEG C of lower seals.Being made into concentration with water-dispersion is 1 × 10
7measure by embodiment 2 after the spore suspension of spore/mL and sprout percentage.
Field efficacy: select Phyllotreta striolata and the simultaneous wild cabbage field of small cabbage moth respectively, being watered by preparation and being dispersed into concentration is 1 × 10
7blade face and ground spraying after the spore suspension of spore/mL, formulation rate 50L/ mu.Adopted parallel great-jump-forward to sample with after medicine 15 days before medicine, 5 points are investigated in every community, and statistics borer population alive, calculates
, repeat 3 times, get its mean value, as the preventive effect of this sample.
The moistening dispersing characteristic of table 3 Metarhizium anisopliae CQMa421 wettable powder, storage period and preventive effect
The results are shown in Table 3.The wettability of test recipe is good, wetting time < 60s; Shelf-stable, stores the active > 80% of spore of 12 months at 28 DEG C; Field is stablized Phyllotreta striolata and small cabbage moth preventive effect, and preventive effect reaches more than 80%, can as the alternative agricultural chemicals of chemical pesticide.
Claims (7)
1. the application of Metarhizium anisopliae CQMa421 bacterial strain in the medicament preparing control Phyllotreta striolata or small cabbage moth.
2. apply as claimed in claim 1, it is characterized in that: the medicament of described control Phyllotreta striolata or small cabbage moth is made up of, all with weight percent green muscardine fungus CQMa421 xerospore powder 44-60%, diatomite 30-40%, nano-aluminum hydroxide 0.5-1%, Xylo-Mucine 3-5%, tween-80 3-5% and OP-10 3-5%.
3. a Metarhizium anisopliae preparation, it is characterized in that: it is made up of, all with weight percent diatomite 30-40%, nano-aluminum hydroxide 0.5-1%, Xylo-Mucine 3-5%, tween-80 3-5%, OP-10 3-5% and green muscardine fungus CQMa421 xerospore powder 44-60%.
4. Metarhizium anisopliae preparation as claimed in claim 3, is characterized in that: described tween-80 and OP-10 are tween-80 and the OP-10 of purity >95%; Described Xylo-Mucine is that purity is greater than 95%; Described diatomite be white to light brown loose powder, purity is greater than 85%, water content is less than 1%; Described nano-aluminum hydroxide is the dewatering nano aluminium hydroxide of silicon fluoride base, and particle diameter is less than 50nm; Described green muscardine fungus CQMa421 xerospore powder is pure spore powder, spore content >=15,000,000,000 spores/gram, water content≤5%; Above all in mass percentage.
5. Metarhizium anisopliae preparation as described in claim 3 or 4, it is characterized in that: it is made up of, all with weight percent diatomite 35-40%, nano-aluminum hydroxide 0.5-0.8%, Xylo-Mucine 3-5%, tween-80 3-5%, OP-10 3-5% and green muscardine fungus CQMa421 xerospore powder 45-55%.
6. as claim 3, Metarhizium anisopliae preparation described in 4 or 5, it is characterized in that: described green muscardine fungus CQMa421 xerospore powder is strong virus force Metarhizium anisopliae CQMa421 spore powder, described strong virus force Metarhizium anisopliae CQMa421 spore powder is obtained by following methods: at 28 DEG C, in humidity 50-70% environment, Metarhizium anisopliae CQMa421 is inoculated in Phyllotreta striolata imago breeding, select the Phyllotreta striolata of susceptible death at first, put into 28 DEG C of incubator moisturizings and cultivate 3-5 days, its insect surfaces is made to grow Fresh spores, collect Fresh spores, inoculation 3-4 diamondback moth larvae in age, the Fresh spores that the same acquisition insect surfaces grows, circulation inoculation Phyllotreta striolata described above and small cabbage moth obtain the Metarhizium anisopliae CQMa421 Fresh spores after taming for 16 times, aseptically, ruling containing on the 1/4SDA culture medium flat plate of 100 mcg/ml penbritins with Fresh spores after the domestication of transfering loop picking, the culture dish of having rule upset is being positioned in 28 DEG C of thermostat containers and cultivates, the typical case grown is selected and without the bacterium colony of miscellaneous bacteria after 3-4 day, at colony edge with moving substratum one fritter of acicula picking with mycelia, proceed to test tube slant substratum central authorities, 28 DEG C of Metarhizium anisopliae CQMa421 mothers obtaining strong virus force for constant temperature culture 10-15 days plant, CQMa421 bacterial strain mother being planted is seeded on 1/4SDA flat board, cultivate 15 days, collect ripe conidium and obtain green muscardine fungus CQMa421 xerospore powder for 28 DEG C.
7. one kind strengthens the method for Metarhizium anisopliae virulence, it is characterized in that: 28 DEG C, in humidity 50-70% environment, Metarhizium anisopliae CQMa421 is inoculated in Phyllotreta striolata imago breeding, select the Phyllotreta striolata of susceptible death at first, put into 28 DEG C of incubator moisturizings and cultivate 3-5 days, make its insect surfaces grow Fresh spores, collect Fresh spores, inoculation 3-4 diamondback moth larvae in age, the Fresh spores that the same acquisition insect surfaces grows; Circulation inoculation Phyllotreta striolata described above and small cabbage moth obtain the Metarhizium anisopliae CQMa421 Fresh spores after taming for 16 times; Aseptically, ruling containing on the 1/4SDA culture medium flat plate of 100 mcg/ml penbritins with Fresh spores after the domestication of transfering loop picking, the culture dish of having rule upset is being positioned in 28 DEG C of thermostat containers and cultivates; The typical case that grows is selected and without the bacterium colony of miscellaneous bacteria after 3-4 days, at colony edge with moving substratum one fritter of acicula picking with mycelia, proceed to test tube slant substratum central authorities, 28 DEG C of Metarhizium anisopliae CQMa421 mothers obtaining strong virus force for constant temperature culture 10-15 days plant.
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| CN106434362A (en) * | 2016-08-11 | 2017-02-22 | 华南农业大学 | Anti-ultraviolet high-toxicity meterhizium anisopliae mutant strain MaUV-1 and application thereof |
| CN108703154A (en) * | 2018-08-07 | 2018-10-26 | 重庆聚立信生物工程有限公司 | Green muscardine fungus kills roach agent and preparation method thereof with boric acid or borax compounding |
| CN110558333A (en) * | 2019-09-24 | 2019-12-13 | 重庆谷百奥生物研究院有限公司 | Composition for killing prodenia litura by metarhizium anisopliae and metaflumizone |
| CN110669675A (en) * | 2019-10-25 | 2020-01-10 | 云南农业大学 | Metarhizium anisopliae MANGS71814 and application thereof in control of potato tuber moth |
| GB2577250A (en) * | 2018-09-14 | 2020-03-25 | Bionema Ltd | Insect-pathogenic fungus, spores, composition and use of same |
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| CN106434362A (en) * | 2016-08-11 | 2017-02-22 | 华南农业大学 | Anti-ultraviolet high-toxicity meterhizium anisopliae mutant strain MaUV-1 and application thereof |
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| GB2577250A (en) * | 2018-09-14 | 2020-03-25 | Bionema Ltd | Insect-pathogenic fungus, spores, composition and use of same |
| CN110558333A (en) * | 2019-09-24 | 2019-12-13 | 重庆谷百奥生物研究院有限公司 | Composition for killing prodenia litura by metarhizium anisopliae and metaflumizone |
| CN110669675A (en) * | 2019-10-25 | 2020-01-10 | 云南农业大学 | Metarhizium anisopliae MANGS71814 and application thereof in control of potato tuber moth |
| CN110669675B (en) * | 2019-10-25 | 2021-07-06 | 云南农业大学 | Metarhizium anisopliae MANGS71814 and application thereof in control of potato tuber moth |
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