CN103783030B - The method of the green ground beetle of a kind of predatory natural enemy insect and green muscardine fungus combination control insect - Google Patents
The method of the green ground beetle of a kind of predatory natural enemy insect and green muscardine fungus combination control insect Download PDFInfo
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Abstract
The present invention relates to field of biological control, specifically, it relates to a kind of method of green ground beetle of predatism natural enemy and green muscardine fungus combination control insect, described method comprises the step thrown in through the inoculated green ground beetle of metarrhizium agent. The present invention proposes a kind of new biological prevention, it is conceived to combine both " the controlling worm with worm " and " controlling worm with bacterium " in biological control, explores a kind of novel controlling way. Playing predatism natural enemy and prey on prevention and control for a long time, green muscardine fungus each formulation advantage and the anti-effect of mixing, prevention effect significantly improves, and uses at any time and prepares at any time, and effective, the time length is long. Save manpower and materials, shuttling back and forth of ground beetle can be utilized to be propagated by microbial pesticide with reactivity, promote species diversity, it is achieved the sustainable control of insect.
Description
Technical field
The present invention relates to field of biological control, specifically, it relates to a kind of method of green ground beetle of predatism natural enemy and green muscardine fungus combination control insect.
Background technology
In recent decades, owing to relying on merely chemical pesticide and abuse chemical pesticide, so that 3R has occurred, not only kill and wound natural enemy, disturbing the Natural control action of natural enemy, and jeopardize the safety of people, biological control is more and more profoundly understood and widespread use by people.
Biological control, refers to the method that harm occurs or reduces utilizing beneficial organism and meta-bolites control harmful organism population thereof. Generally refer to and control worm, fungus treatment with worm, cure the disease with bacterium, control the methods such as grass, transgenic breeding, property telergone with bacterium. Biological pesticide refers to and utilizes living organisms (fungi, bacterium, insect viruses, genetically modified organism, natural enemy etc.) or its meta-bolites to carry out the preparation killed or suppress for agricultural pest. Biological pesticide has free from environmental pollution, people and other biological safety, preventive and therapeutic effect is lasting, product noresidue, sick worm is killed and wounded high specificity; it is easy to same other plant sfgd. cooperation and the advantages such as energy save energy, more and more important effect will be played in integrated pest management.
Green muscardine fungus belongs to Mycophyta (Eumycota) Ascomycotina (Ascomycotina) gang pyrenomycetes (Pyren-mycetes) Spheeriales (Sphaeriales) Clavicipitaceae (Clavisipitetaceae), it it is the important biocontrol fungi of a class, can parasitic various insects, production is widely used, according to incompletely statistics, the whole world about has 200 various insects can be lethal by this kind of fungi infestation. This bacterium virulence is strong, effective, to people, animal and farm crop toxicological harmless, is that on our times, one of maximum entomogenous fungi is applied in research.
Fungi biological prevention and control agent has tagging property, popularity, the environmentally safe and feature such as not easily develop immunity to drugs. Utilizing entomogenous fungi control insect to become an important aspect of the comprehensive regulation based on biological control, entomogenous fungi is the important factor of pest population Natural regulation and the important materials of biological control of insect pests. At present, the green muscardine fungus preparation about more than 10 of research and development is planted both at home and abroad, wherein, affect maximum and the most successful example be Britain CABIBio2science locust biological control international cooperative project (LUBILOSA) develop kill locust green muscardine fungus biological pesticide, in Africa registration, go into operation, big area is used for the control of desert locust, becomes effective locust biological prevention and control agent.
Green ground beetle Carabus (Coptolabrus) smaragdinusFischervonWaldheim is subordinate to Coleoptera Coleoptera, ground beetle section Carabidae, ground beetle race Carabini, ground beetle belongs to Carabus, it it is a kind of predatory natural enemy insect resource having important utility value, it is distributed in major part province, the north at home, it is distributed in the southeast in siberian, Far-east Area of Russia and Korea S abroad. The predatism life of this worm battalion, is mainly food taking earthworm, is played an important role by maintaining ecological balance. Green ground beetle is the important predatism natural enemy in farmland, and it becomes, larva prey ability is strong, Quick off the mark, and special eating snail, lepidopterous insects, also take food grub, locust etc., utilizes this ground beetle control agricultural pests to have broad application prospects
But it is low that at present biological control also exists anti-effect, resource dispersion, to prevent and treat resource single, the problems such as prevention and control mode is single. Therefore integration of innovational resources, the mode of action of innovation biological control, it is to increase the anti-effect of biological control is biological control urgent problem. Green muscardine fungus has stronger parasitic specificity, the green ground beetle of predatory natural enemy insect, logical edge ground beetle, black ground beetle etc. is nontoxic, therefore, predatism natural enemy stronger for activity ability and entomogenous fungi are combined, the function of predatism natural enemy predation insect can either be played, natural enemy insect can be utilized again on a large scale to carry muscardine spore powder and propagate, saving manpower and materials, increase the probability of insect green stiff fungus infection, cause the epiphytotics generation of green muscardine fungus. Organically combining Control pests by predatism natural enemy and green muscardine fungus two kinds of biological pesticides, the associating effect playing both has very wide application prospect.
Summary of the invention
It is an object of the invention to provide a kind of green ground beetle of predatism natural enemy and the method for green muscardine fungus combination control insect.
The present invention utilizes the green ground beetle entomogenous fungi green muscardine fungus combination of predatory natural enemy insect, plays the advantage of each form biological pesticide, it is to increase the scope of control harmful organism and anti-effect, the biological pesticide of the safety non-pollution of effective Control pests.
The green ground beetle of predatism natural enemy and the method for green muscardine fungus combination control insect according to the present invention comprise the following steps:
(1) muscardine spore powder is prepared;
(2) green to green muscardine fungus and predatism ground beetle is combined;
(3) to be stained with the ground beetle invasion and attack insect of spore powder.
Method according to the present invention, preparation green muscardine fungus 107��108Individual spore/ml, wettable powder green muscardine fungus 100-200 hundred million spore/gram.
According to the specific embodiment of the present invention, being combined by green to green muscardine fungus and predatism ground beetle, step is as follows:
(1) plastics casing of an about 1m*0.7 that can hold ground beetle is prepared;
(2) green muscardine fungus wettable powder is sprinkling upon in big box, spore powder thickness about 0.2cm;
(3) ground beetle is put into wherein;
(4) suspended emulsion agent is diluted 100 times, with box sizableness, the gauze of thickness about 1cm, gauze dilute after suspended emulsion agent soak;
(5) being laid in the box filling ground beetle by the gauze being stained with muscardine spore powder, ground beetle is covered by gauze, waiting 5-10 minute can take off gauze, and the ground beetle being stained with spore powder in box can discharge, for preventing and treating insect.
Present method can be used for according to the difference of green muscardine fungus preparation and the feeding habits of green ground beetle preventing and treating migratory locusts, grassland grasshopper, fall webworms, snail, slug, white star brevitarsis, loxostege sticticalis, black cutworm, big cutworm, apple tip hawk noctuid, Phalera flavescens, white tiger moth, diaphania, mulberry geometrid (Hemerophila atrilineata), gypsymoth etc.
It is low that current biological control also exists anti-effect, and resource dispersion, to prevent and treat resource single, the problems such as prevention and control mode is single. Therefore integration of innovational resources, the mode of action of innovation biological control, it is to increase the anti-effect of biological control is biological control urgent problem. The present invention proposes a kind of new biological prevention, it is conceived to combine both " the controlling worm with worm " and " controlling worm with bacterium " in biological control, explores a kind of novel controlling way.
The advantage of the present invention:
1. predatism ground beetle has taking food property of wide spectrum and can control various pests, and predation and activity ability are relatively strong, and scope of activity is wider.
2. microbial pesticide derives from physical environment, the single-minded parasitics of tool.
3. the biology such as pair people, animal is safer, and environmentally safe is nuisanceless, green, safety.
4. pair natural enemy is safer, and it has the selectivity to target pest poisoning, it is not easy to develop immunity to drugs.
5. production technique is fairly simple, and the expenses such as exploitation and registration are lower than characteristics such as chemical pesticides.
6. playing predatism natural enemy and prey on prevention and control for a long time, green muscardine fungus each formulation advantage and the anti-effect of mixing, prevention effect significantly improves
7. using at any time and prepare at any time, effective, the time length is long.
8. save manpower and materials, shuttling back and forth of ground beetle can be utilized to be propagated by microbial pesticide with reactivity, promote species diversity, it is achieved the sustainable control of insect.
Accompanying drawing explanation
Fig. 1 is corrected mortality graphic representation.
Fig. 2 shows green muscardine fungus process to the impact of green ground beetle adult mortality ratio.
Fig. 3 shows green muscardine fungus and green ground beetle and different combined treatment to the lethal power of Asiatic migrotory locust, wherein, and I: the green ground beetle of release separately; II: use separately green muscardine fungus; III: discharge green ground beetle and use green muscardine fungus simultaneously; IV: release after green ground beetle inoculation green muscardine fungus.
Embodiment
Embodiment 1
1.1 test materials
The nymph of a locust in 3 ages: pick up from Tai'an, Shandong plant protection institute of Shandong Agricultural University Asiatic migrotory locust breeding base. Kill the female powder LS20041884 of locust muscardine spore: Chongqing fungal farm chemicals is developed engineering center and provided. With 0.2% tween solution, Metarhizium anisopliae spore powder is diluted to 105Individual/mL, 106Individual/mL, 107Individual/mL, 108Individual/mL, for subsequent use. Green ground beetle derives from Shandong Agricultural University's plant protection institute environmental organism and insect resources institute, the green ground beetle raised with Asiatic migrotory locust, tames through the Asiatic migrotory locust predatory feeding of a year.
1.2 Metarhizium anisopliaes are to Asiatic migrotory locust toxicity test
Adopt leaching poison method process examination worm, draw the muscardine spore liquid of 2uL with pipettor, take out after nymph of a locust back is immersed muscardine spore liquid, 15, every cage. 0.2% tween solution of control group 2uL. Each group of process repeats 3 times.After inoculation, examination worm being placed on T=(28 �� 1 DEG C), the environment pilot scale that humidity is 50%��70%, 24h illumination is supported. 7:30 fed once new leaf of Semen Maydis and removed foreign material every morning, recorded the quantity of nymph of a locust death simultaneously, and continuous observation is until the corrected mortality of each group reaches more than 80%. The locust checkmated from the 3rd day is taken out and carries out moisturizing (culture dish is spread the filter paper soaked with sterilized water or put the absorbent cotton soaked, and each locust corpse can not contact to some extent, finally records the stiff worm rate of locust).
Dosage of inoculation (individual/g)=[inoculum density (individual/mL) �� inoculation volume (mL)]/examination polypide is heavy
Corrected mortality: M=(t c)/(100 c) �� 100%
M: corrected mortality c: comparison mortality ratio t: process mortality ratio
The regression equation of statistic between each process is detected by variance analysis, Deng's Ken Shi duncan's new multiple range method.
Raw result of surveying shows, and a few days ago all dead without the nymph of a locust, from the 3rd day, spore concentration was 10 �� 107(individual/mL) and 1.0 �� 108(individual/mL) has the nymph of a locust dead, and corrected mortality is respectively 4.67% and 11.13%. Along with the increase of number of days, the mortality ratio of polypide is also in increase. When the 10th day, spore concentration is 1.0 �� 108The treatment group of (individual/mL), corrected mortality reaches 100%, and now, spore concentration is 1.0 �� 1.07Individual/mL, 1.0 �� 106Individual/mL, 1.0 �� 105The treatment group of individual/mL, corrected mortality is respectively 93.33%, and 75.57%, 64.46%. When the 14th day, the corrected mortality of all treatment group is all 100%.
From table 1 Metarhizium anisopliae to 3 age Asiatic migrotory locust the result of virulence it may be seen that on the conspicuous level of degree of confidence 95%, spore concentration is 1.0 �� 106Individual/mL, 1.0 �� 105Individual/mL to 3 age Asiatic migrotory locust the difference of virulence very remarkable, and 1.0 �� 1.07Individual/mL, 1.0 �� 108Individual/mL to 3 age Asiatic migrotory locust the difference of virulence not obvious. Totally, it seems, LD50Size order be 1.0 �� 108Individual/mL < 1.0 �� 107Individual/mL < 1.0 �� 106Individual/mL < 1.0 �� 105Individual/mL, wherein, the spore concentration of best results is 1.0 �� 108Individual/mL.
Table 1 processes the corrected mortality (%) of the rear nymph of a locust
Table 2 Metarhizium anisopliae to 3 age Asiatic migrotory locust virulence
Note: with letter representation significant difference different after column data, Duncans method, P < 0.05
1.3. green muscardine fungus is to green ground beetle safety evaluation
Getting 0.1 muscardine spore powder makes its surrounding be uniformly coated on 1m �� 0.7m plastics casing, makes plastics casing cover last layer bacterium powder. 50 green ground beetles are put into box tenebrio molitor forage feed, records green ground beetle death condition every day. Its moisturizing being processed after green ground beetle death, whether inspection has stiff worm to occur. If not executing bacterium powder for comparison, often process 3 repetitions.
Observed and recorded is through green muscardine fungus process with without the mortality ratio of green ground beetle every day of green muscardine fungus process and stiff worm rate, green ground beetle after life 36d except part is dead, s-generation larva occurs and activity is good, other all enter more winter condition, and treatment group and control group survivorship curve coincide (Fig. 2) substantially. The s-generation larva when 27 days occurs control group, and the green ground beetle of green muscardine fungus process s-generation larva also occurred when 33 days; Variance analysis, probability P r is carried out between process > F, illustrate that green muscardine fungus treatment group and comparison are without significant difference. The green muscardine fungus of death is carried out moisturizing process, after 10d, does not all find the growth of green muscardine fungus mycelia, illustrate that green muscardine fungus is to green ground beetle adult safety.
1.4 green muscardine funguss are combined the control action kou to Asiatic migrotory locust with green ground beetle
Field modeling green muscardine fungus and green ground beetle different composite form Asiatic migrotory locust lethal effect, use 1m3Shroud field feed 3 age Asiatic migrotory locust, by green ground beetle Nature enemy 1 day before test. The process green ground beetle of I(is to the lethal power of Asiatic migrotory locust): choose female, male each one of healthy green ground beetle, put into shrouding containing 10 3 Asiatic migrotory locusts in age; Process II: by concentration 1 �� 108Above the migratory locusts that individual/mL green muscardine fungus bacterium agent 2ul is sprayed onto in shrouding and food thereof, observed and recorded green muscardine fungus is to the parasitic lethality rate of Asiatic migrotory locust (Asiatic migrotory locusts in 3 ages); Process III: throw in healthy ground beetle female, male each one, spray green muscardine fungus bacterium agent simultaneously, the green ground beetle of observed and recorded to the ingestion rate of Asiatic migrotory locust and green muscardine fungus to the parasitic lethality rate of Asiatic migrotory locust (Asiatic migrotory locusts in 3 ages); Process IV: throwing in through the inoculated green ground beetle of metarrhizium agent, the green ground beetle after observed and recorded entomogenous fungi compound is to the compound lethality rate of Asiatic migrotory locust (Asiatic migrotory locusts in 3 ages). CK: spray 2ul sterilized water to do not put green ground beetle and green muscardine fungus containing in the shrouding of 10 Asiatic migrotory locusts in contrast, each process repeats three times, every day, foodstuff was changed in timing, add up the lethality rate of Asiatic migrotory locust every day respectively, and the Asiatic migrotory locust having green muscardine fungus to process is taken out and carries out moisturizing, observe and calculate stiff worm rate.
Asiatic migrotory locust is simulated field test and shows by green muscardine fungus combination different from green ground beetle; the correction lethality rate of Asiatic migrotory locust is reached 83.33%, 100% from 2 kinds of different combination treatment methods (process III, process IV) of green ground beetle by green muscardine fungus respectively, is all significantly higher than the 83.33% of 66.67% and green ground beetle single dose treatment group that green muscardine fungus single dose processes; But the significance of difference between 2 kinds of different array modes, it may be that owing to the interaction relationship between green ground beetle and green muscardine fungus with Asiatic migrotory locust three is relevant, causes singly granting complex role mode different from action function. Simultaneously, it may also be useful to green muscardine fungus and green ground beetle and the process III that both do not combined, green ground beetle ingestion rate declines to some extent than process I and process IV. Dong Hui (2005) proves that ground beetle does not take food by the locust of green muscardine fungus parasitism, and therefore this may identify by the Asiatic migrotory locust of parasitism due to green ground beetle, thus reduces its predation. And the green muscardine fungus infection rate processing III rises than processing I and process IV, this may decline due to the Asiatic migrotory locust vitality frightened by the predation of green ground beetle or baited, and injured Asiatic migrotory locust is more easily infected; Meanwhile, green ground beetle, as the communication media of green muscardine fungus, adds spread scope and the probability of green muscardine fungus, also makes Asiatic migrotory locust more easily be subject to infecting to green muscardine fungus.
Test also shows, the success ratio that takes food of locust is increased and decline age by green ground beetle with the worm of locust. After 14d, locust major part is sprouted wings, and locust number of individuals on average remains unchanged for continuous 4.2 days. Comprehensive test result, by green muscardine fungus and green ground beetle compound use that the control action kou effect of Asiatic migrotory locust is best, this kind of composite mode of combination, taking green ground beetle as communication media had both saved human and material resources, financial resources, play again advantage and the function of each form biological pesticide, improve the control scope of harmful organism and anti-effect, the breaking out and endanger of effective Control pests of safety non-pollution.
Comparison between table 3 different treatment Asiatic migrotory locust ingestion rate, infection rate and lethality rate
Note: with letter representation significant difference different after column data, Duncans method, P < 0.05
Although, prior art has report, when utilizing pathogenic fungi control insect, can endanger natural enemy, but, instant invention overcomes the problems referred to above, as long as filtering out suitable fungal pathogen, risk being dropped to minimum degree.The green muscardine fungus purifying of acquisition is diluted to different concentration by the present invention, then, using the green muscardine fungus after the Asiatic migrotory locust of artificial breeding and green ground beetle and dilution as test materials, infusion method is adopted to be inoculated into by green muscardine fungus with it green ground beetle, carry out toxicity test, it is safe for demonstrating green muscardine fungus to green ground beetle, and two kinds of biological and ecological methods to prevent plant disease, pests, and erosion methods can use simultaneously. Then carry out field modeling test, by observing green muscardine fungus to the parasitism of Asiatic migrotory locust and green ground beetle to activities such as the predations of Asiatic migrotory locust, have studied green muscardine fungus and the effect of Asiatic migrotory locust prevented and treated jointly by green ground beetle. Use green ground beetle to spray green muscardine fungus process simultaneously and throw in through metarrhizium agent inoculated green ground beetle, the control effects of Asiatic migrotory locust to be all significantly higher than green muscardine fungus single dose and infect and prey on separately with green ground beetle, and the inoculated green ground beetle of metarrhizium agent is best to Asiatic migrotory locust control effects.
The green muscardine fungus that the present invention adopts and the combination of predatism ground beetle also have good effect for control lepidoptera pest such as noctuid, small heart-eating peach worm, oriental fruit months etc., by changing combination green muscardine fungus preparation used and other large-scale predatism ground beetles, it is possible to for prevention and control and the improvement of more harmful organisms.
The present invention proves that natural enemy and pathogenic micro-organism can act synergistically and controls harmful insect and have bigger synergism, predatism green ground beetle build is big, there is activity ability strong, the advantages such as predation insect is wide, some ground beetle energy Grasshopper, carry spore convenient, disseminate green muscardine fungus bacterium spore scope and efficiency height, so, predatism natural enemy stronger for activity ability and entomogenous fungi are combined, the function of predatism natural enemy predation insect can either be played, natural enemy insect can be utilized again on a large scale to carry muscardine spore powder and propagate, save manpower and materials, increase the probability of insect green stiff fungus infection, cause the epiphytotics generation of green muscardine fungus. therefore, organically combining Control pests by green for predatism natural enemy ground beetle and green muscardine fungus two kinds of biological pesticides, the associating effect playing both has very wide application prospect.
Embodiment 2
Predatism walks green ground beetle mass rearing and breeding, and chooses the adult that violent activity ability is stronger, female-male proportion 1:1.
Obtain fermenting green muscardine fungus spore powder, it is processed into wettable powder: with the former medicine of green muscardine fungus, inert filler and a certain amount of auxiliary agent, in proportion through abundant co-grinding, reaches certain particle size and make wettable powder. Through experiment research and bioassay, the best muscardine spore wettable powder being applicable to meets:
1. content, green muscardine fungus active constituent content 100-200 hundred million spores/g; 2. suspensibility 93%; 3. wetting time: wettability 56 seconds. 4. potential of hydrogen (pH value) PH=7.8. 5. moisture: moisture content is less than 3%. muscardine spore wettable powder formulas and is: be weight, 60% spore, 30% weighting agent, 8% blender, 2% wetting agent.
Suspended emulsion agent is prepared: muscardine spore, salad oil are made into emulsion in proportion; and it is active to add xitix protection spore; add SDS and finally make suspended emulsion agent as emulsifying agent; through experiment research and test; the best muscardine spore suspended emulsion agent proportioning being applicable to is: spore/salad oil ratio 15%; 0.2% xitix, 0.1SDS.
Directly being sprinkling upon in plastics casing by wettable powder, thickness reaches 0.1-0.3cm, is put in a large number by ground beetle, often large puts into ground beetle 150 about-200
Suspended emulsion agent thickness being about the dilution of 1cm gauze soaks, and above ground beetle gauze being layered in box, contact ground beetle, allows the ground beetle back side be stained with spore.
Waiting 5-15 minute, ground beetle polypide speckles with muscardine spore, then can be discharged into field.
Claims (4)
1. the method for the green ground beetle of predatism natural enemy and green muscardine fungus combination control insect, it is characterised in that, described method comprises the step thrown in through the inoculated green ground beetle of metarrhizium agent.
2. the method for the green ground beetle of predatism natural enemy according to claim 1 and green muscardine fungus combination control insect, it is characterised in that, described insect is locust, noctuid, small heart-eating peach worm or oriental fruit months.
3. the method for the green ground beetle of predatism natural enemy according to claim 1 and green muscardine fungus combination control insect, it is characterised in that, described metarrhizium agent is muscardine spore powder, and its concentration is 107��108Individual spore/ml.
4. the method for the green ground beetle of predatism natural enemy according to claim 1 and green muscardine fungus combination control insect, it is characterized in that, described metarrhizium agent is the wettable powder green muscardine fungus that muscardine spore powder is processed into, its green muscardine fungus active constituent content 100-200 hundred million spores/g.
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