CN105123766A - Entomopathogenic nematode HbSD and insecticide thereof, preparation method and application - Google Patents
Entomopathogenic nematode HbSD and insecticide thereof, preparation method and application Download PDFInfo
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- CN105123766A CN105123766A CN201510653569.2A CN201510653569A CN105123766A CN 105123766 A CN105123766 A CN 105123766A CN 201510653569 A CN201510653569 A CN 201510653569A CN 105123766 A CN105123766 A CN 105123766A
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Abstract
The invention discloses an entomopathogenic nematode HbSD and insecticide thereof, a preparation method and an application. The preparation method of a suspension liquid preparation comprises the steps that last instar flour weevil larvas are infected with the HbSD, red dead insects are obtained on the sixth day, the HbSD continues to be cultured to the tenth day, the HbSD crawls out of dead insect corpses into water, and suspension liquid is obtained; Triton X-100 is added to the suspension liquid, and the entomopathogenic nematode HbSD suspension liquid preparation is prepared. The preparation method of an insect corpse agent comprises the steps that last instar flour weevil larvas are infected with the HbSD, reddish brown dead insects are obtained on the sixth day, and the HbSD continues to be cultured to the ninth day to obtain the insect corpse agent. The suspension liquid preparation and the insect corpse agent have the advantages of being stable in effect, easy to produce, low in cost, free of pollution, high in injurious insect killing activity and the like.
Description
Technical field
The present invention relates to biological insecticides, particularly relate to entomopathogenic nematode HbSD, its insecticide and preparation method and application.
Background technology
Entomopathogenic nematode is the entomogenous nematode carrying symbiotic bacteria in a class body, and such nematode, as biological insecticides novel in the world, has host range extensive; To vertebrate, plant and other non-target organism safety, be beneficial to environmental protection; Be easy to carry out live body or in vitro mass propgation, easy to use; Initiatively can find host, kill off the insect pests fast; Can recycle in the environment, can with the advantage such as number of chemical is mixed with biological insecticides, in the multiple soil pests of control and hidden insect, played important effect.
Entomopathogenic nematode is under the jurisdiction of animal kingdom's Nemathelminthes (Nematoda), phasmid (Secernentea), Rhabditida (Rhabditida), up to now, the known nematode parasitizing insect is about 40 Duo Ge sections, more desinsection nematode is wherein used to comprise Steinernema Carpocapsae (Steinernemacarpocapsae, Sc) and Heterorhabditis bacteriophora-NJ (Heterorhabditisbacteriophora, Hb).The bacterium of nematode and its parachorium acts synergistically to kill insect.Nematode for desinsection is considered to the biological insecticides to environment and mankind close friend.Current desinsection nematode has been widely used in dwelling property of soil insect, as chafer, wireworm, cutworm, mole cricket, root maggot, and the control of the insects such as leek maggot.Application process comprises the worm corpse after nematode spray suspension liquid and nematode infections; but because its spray suspension liquid need add 4-5 kind auxiliary agent and protectant (as anti-evaporating agent, water-loss reducer, adhesive, uv-protector); cause its production cost high, and its application of disadvantages affect such as prepared insecticide insecticide efficiency is low.
Summary of the invention
A kind of entomopathogenic nematode HbSD provided by the invention, improves insecticide efficiency.
On the other hand, suspension preparation provided by the invention solves biological insecticides production process need increase multiple auxiliary agent and protectant causes high in cost of production problem.
Entomopathogenic nematode HbSD of the present invention, by being placed in nematode by children yellow mealworm in age, making end yellow mealworm in age become the dead worm of rufous, filtering out nematode HbSD further behind some skies.
The preparation method of entomopathogenic nematode HbSD suspension preparation, comprise and HbSD is infected end Yellow meal worm larva in age, within 6th day, obtain the dead worm that color becomes rufous, wet filter paper continues be cultured to the 10th flying worm HbSD infective stage larva climb out of in dead worm corpse, enter in water, obtain suspension, remain on 10-12 DEG C; Add TritonX-100 toward suspension during use and obtain HbSD suspension preparation.
Entomopathogenic nematode HbSD suspension preparation, comprises entomopathogenic nematode HbSD and TritonX-100.
Entomopathogenic nematode HbSD suspension preparation, the entomopathogenic nematode HbSD stoste being 900-1000 bar/mL by concentration during use is made into 27-30 bar/mL entomopathogenic nematode HbSD and volume accounting is the TritonX-100 of 0.05%.
The preparation method of entomopathogenic nematode HbSD worm corpse agent, uses entomopathogenic nematode HbSD to infect end Yellow meal worm larva in age, within the 6th day, obtains the dead worm that color becomes rufous, continue to be cultured to the 9th day and obtain worm corpse.
Entomopathogenic nematode HbSD suspension preparation for preventing and treating insect, insect be end age greater wax moth larva, 3 age Sugarcane Yellow snout moth's larva larva, end age Yellow meal worm larva, Brontispa longissima larva or end age Clania variegata Snellen larva.
The agent of entomopathogenic nematode HbSD worm corpse for preventing and treating insect, insect be end age greater wax moth larva, 3 age Sugarcane Yellow snout moth's larva larva, end age Yellow meal worm larva, end age Brontispa longissima larva or end age Clania variegata Snellen larva.。
The present invention obtains entomopathogenic nematode HbSD by screening, and finds that it has good effect to pest control.
Further, nematode suspension preparation of the present invention and the agent of worm corpse.These two kinds of preparations are as insecticide, and preparation method and application's method is all simpler.Compared with prior art, the preparation and application of nematode killer is simplified.
Entomopathogenic nematode HbSD suspension preparation of the present invention and the agent of worm corpse all have effect stability, produce simple and easy, with low cost, pollution-free, bactericidal activity advantages of higher.
Accompanying drawing explanation
Fig. 1 is the pesticidal experimental result pictures of 4 kinds of mass production nematodes to end greater wax moth larva in age;
Fig. 2 be entomopathogenic nematode HbSD infective stage larva respectively to 3 age Sugarcane Yellow snout moth's larva larva and end age Yellow meal worm larva pesticidal experimental result picture;
Fig. 3 is that the corpse agent of entomopathogenic nematode HbSD worm and entomopathogenic nematode HbSD suspension preparation are respectively to the control efficiency figure of Sugarcane Yellow snout moth's larva;
Fig. 4 is the experimental result picture that garden sprays that entomopathogenic nematode HbSD suspension preparation prevents and treats fragrant camphor tree Clania variegata Snellen insect.
Embodiment
By reference to the accompanying drawings the present invention is described in further detail below by embodiment.
(Hb is that heterorhabditis indica is called for short to entomopathogenic nematode HbSD of the present invention, SD is numbering) be a kind of heterorhabditis indica, by the side's of trapping acquisition in the wild, concrete grammar is: be placed in nylon wire by end Yellow meal worm larva in age, imbed in Field Soil, within 6th day, results color becomes the yellow mealworm of rufous, and the nematode parasitized in yellow mealworm is entomopathogenic nematode HbSD of the present invention.HbSD has good fecundity, also can obtain its offspring by artificial mass propgation.
Entomopathogenic nematode HbSD infective stage larva (IJs) is the insect larvae after infected insect pathogenic nematode HbSD.
Embodiment one: the preparation method of entomopathogenic nematode HbSD suspension preparation
The first entomopathogenic nematode HbSD insecticide is suspension preparation, and its preparation method comprises:
The preparation method of entomopathogenic nematode HbSD suspension preparation: put one deck filter paper at plastic box bottom, the Yellow meal worm larva 100 in last age selecting healthy growth is placed on filter paper, be added on the yellow mealworm in age of end with the HbSD nematode hanging drop that liquid-transfering gun absorption concentration is 1000/ml, cover, polypide color situation of change is checked: the 6th day after room temperature places 4-5 days, the yellow mealworm color of normal infection entomopathogenic nematode becomes rufous, continue to cultivate after discarding off-colour yellow mealworm, within the 10th day, results have infected the yellow mealworm of entomopathogenic nematode HbSD.By " white trapping " method, namely the hydrotaxis of HbSD infective stage larva (IJs) is utilized to collect nematode, nematode crawls into water in insect bodies, discarding of first day results, the infective stage larva suspension of 2-3 days is checked survival rate under the microscope, and the survival rate of nematode is greater than 95%; Then deposit in 10-15 DEG C of refrigerator to connect for next step experiment.
Before using, the nematode suspension concentration of storage is diluted to 900-1000 bar nematode/milliliter, is diluted to the entomopathogenic nematode HbSD of 27-30 bar/mL, adds TritonX-100 in the ratio of liquor capacity 0.05%.
TritonX-100 is Triton X-100, is a kind of nonionic surface active agent, has dispersed and effect that is thickness.
The present embodiment selects TritonX-100 as the single auxiliary agent of nematode HbSD suspension, and cost is low, insecticide efficiency is high and do not affect nematode and find parasitic ability.
Embodiment two: the preparation of entomopathogenic nematode HbSD worm corpse agent
The second entomopathogenic nematode HbSD insecticide is the agent of worm corpse, and its preparation method comprises:
The preparation method of entomopathogenic nematode HbSD worm corpse agent: infect end Yellow meal worm larva in age by with the entomopathogenic nematode HbSD infective stage larva of results on the 1st day.The yellow mealworm selecting color to become rufous on the 6th day, continues to cultivate, and the 9th day results worm corpse, is placed in 50-100mL centrifuge tube, saves backup under 10-14 DEG C of condition.
The survival rate of entomopathogenic nematode HbSD is greater than 95%, and the agent of worm corpse is HbSD metainfective end Yellow meal worm larva in age, and each worm corpse is at least containing 5000 entomopathogenic nematode HbSD.
Two kinds of HbSD nematode killer of the present invention: nematode suspension preparation and the agent of worm corpse.Preparation method and application's method is all simpler.Compared with prior art, the preparation and application of nematode killer is simplified.
Embodiment three: the insecticidal properties experiment of entomopathogenic nematode HbSD
For examination nematode strain: 4 kinds of Heterorhabditis bacteriophora-NJ belong to nematode HbSD, HbI, HbII and HbIII.
For examination insect: end greater wax moth larva in age
Method of testing: indoor bioassay method, namely measures the virulence of 4 kinds of mass production nematode suspension to end greater wax moth larva in age, adds up lethality after 6 days.Control group does not add nematode.
Results and analysis: 4 kinds of mass production nematodes to the pesticidal of end greater wax moth larva in age as shown in Figure 1, at four kinds in examination nematode strain (HbSD, HbI, HbII, HbIII), the insecticidal power of entomopathogenic nematode HbSD of the present invention is the highest, and fecundity is strong.HbSD is selected to be used for further toxicity test.
Embodiment four:
For examination nematode strain: entomopathogenic nematode HbSD infective stage larva
For examination insect: Sugarcane Yellow snout moth's larva (Tetramoeraschistaceana) 3 instar larvae and yellow mealworm linal-instar larvae
Indoor bioassay method, respectively by 3 age sugarcane borer larva or the number ratio of yellow mealworm linal-instar larvae and entomopathogenic nematode HbSD infective stage larva be that 1:100 puts into culture dish, respectively add 200ul water treatment, each process 36 sugarcane borer 3 instar larvaes or yellow mealworm linal-instar larvae, at room temperature place, control experiment, not add entomopathogenic nematode HbSD infective stage larva, only adds running water, adds up lethality after 6 days.
As shown in Figure 2, experimental result shows that entomopathogenic nematode HbSD all has significant virulence, the LC of Sugarcane Yellow snout moth's larva 3 instar larvae to Sugarcane Yellow snout moth's larva 3 instar larvae and end Yellow meal worm larva in age
50value is the LC of 9.00/head, yellow mealworm linal-instar larvae
50value is 16.80/head.
Embodiment five:
For examination material: entomopathogenic nematode HbSD infective stage larva;
For examination insect: greater wax moth (Galleriamellonella.) linal-instar larvae, Brontispa longissima (Brontispalongissim) linal-instar larvae, Clania variegata Snellen (Cryptotheleavariegate) linal-instar larvae.Greater wax moth is cultivated, with fresh coconut leaf feeding Brontispa longissima with artificial feed; From leaf of Cortex cinnamomi camphorae feeding Clania variegata Snellen larva, be all placed in 25 DEG C of climatic cabinate cultivation.
The ratio of all insect infection entomopathogenic nematode infective stage larvas is the same, insect and pathogenic nematode infective stage larva is added bottom and is covered with bottom the culture dish of the graceful filter paper of shellfish, for examination insect Clania variegata Snellen 200, and Brontispa longissima 300, greater wax moth 400.Each process repetition 4 times, recorded lethality after 6 days.
The virulence of HbSD infective stage larva to Brontispa longissima linal-instar larvae, Clania variegata Snellen linal-instar larvae, greater wax moth linal-instar larvae is as shown in table 1, with 8.6-11.7 HbSD head/mL dosage, can kill the Brontispa longissima of 50%, Clania variegata Snellen, greater wax moth linal-instar larvae.
Table 1
Embodiment six:
For examination material: embodiment 1 prepares the entomopathogenic nematode HbSD worm corpse agent of entomopathogenic nematode HbSD suspension preparation and embodiment two preparation
For examination field: there is the sugarcane field that yellow snout moth's larva endangers
Adopt the spray method of suspension preparation, suspension spray amount is 1.02 × 10
4bar/m
2, within one week, spray once, continuous three weeks, each plot area 4m
2, the buffer strip that minizone is 5 meters, each process 6 replicated plots.
The infective stage larva in the worm corpse of embodiment two is counted, about containing 7.71 × 10 in every cephalont corpse under anatomical lens
4entomopathogenic nematode HbSD infective stage larva, worm corpse landfill is the lower 5-8 centimetre of depths of soil around sugarcane plant, hole, 85 centimetres, interval, landfill 2 cephalont corpse in each hole, 6 repetitions.Not add worm corpse for control group, within 30 days, distinguish investigation records result afterwards.Each heatable adobe sleeping platform (1 meter, interval) buries 2 HbSD nematode infections phase worm corpse, can reduce landfill number of times, improves insecticide efficiency, reaches the object of pest control.
Experimental result as Fig. 3 shows, HbSD worm corpse agent (the yellow mealworm corpse of HbSD nematode infection) and HbSD suspension preparation are to the control of Sugarcane Yellow snout moth's larva.After application HbSD suspension preparation process 30d, the relative survival rate of Sugarcane Yellow snout moth's larva larva is reduced to less than 30% than control group.After worm corpse agent process 30d, the relative survival rate of Sugarcane Yellow snout moth's larva larva is reduced to less than 61% than control group.Application HbSD suspension preparation and the agent of HbSD worm corpse all have good control efficiency to Sugarcane Yellow snout moth's larva.
Embodiment seven:
For examination material: entomopathogenic nematode HbSD suspension preparation prepared by embodiment one;
Test field: the area that fragrant camphor tree Clania variegata Snellen morbidity is more serious
The entomopathogenic nematode HbSD suspension preparation using embodiment one to prepare carries out blade back to the more serious fragrant camphor tree of morbidity and sprays; Within every 15 days, spray once, totally 3 times, within 3 months, " Invest, Then Investigate " prevents and treats result.
Garden sprays HbSD suspension preparation and prevents and treats fragrant camphor tree Clania variegata Snellen insect result as shown in Figure 4, and garden sprays HbSD suspension preparation can prevent and treat the outburst of fragrant camphor tree Clania variegata Snellen insect for 3 times effectively.
Above content is in conjunction with concrete embodiment further description made for the present invention, can not assert that specific embodiment of the invention is confined to these explanations.For general technical staff of the technical field of the invention, without departing from the inventive concept of the premise, some simple deduction or replace can also be made.
Claims (9)
1. entomopathogenic nematode HbSD, is characterized in that: described entomopathogenic nematode HbSD infects end Yellow meal worm larva in age, makes end Yellow meal worm larva in age become the dead worm of rufous.
2. entomopathogenic nematode HbSD suspension preparation, is characterized in that: comprise entomopathogenic nematode HbSD and TritonX-100 according to claim 1.
3. entomopathogenic nematode HbSD suspension preparation according to claim 2, is characterized in that: comprise entomopathogenic nematode HbSD that concentration is 27-30 bar/mL and volume accounting is the TritonX-100 of 0.05%.
4. the preparation method of the entomopathogenic nematode HbSD suspension preparation according to Claims 2 or 3, it is characterized in that: infect end Yellow meal worm larva in age with entomopathogenic nematode HbSD, within 6th day, obtain the dead worm that color becomes rufous, continue to be cultured to the 10th day, HbSD climbs out of and enters water in dead worm corpse, obtains suspension; Add TritonX-100 toward suspension and obtain entomopathogenic nematode HbSD suspension preparation.
5. the preparation method of entomopathogenic nematode HbSD worm corpse agent, is characterized in that: adopt entomopathogenic nematode HbSD according to claim 1 to infect end Yellow meal worm larva in age, within the 6th day, obtain the dead worm that color becomes rufous, continue to be cultured to the 9th day and obtain the agent of worm corpse.
6. entomopathogenic nematode HbSD worm corpse agent, is characterized in that: adopt the preparation method of entomopathogenic nematode HbSD worm corpse according to claim 5 agent to obtain.
7. entomopathogenic nematode HbSD worm corpse according to claim 6 agent, is characterized in that: at least contain 5000 entomopathogenic nematode HbSD in each worm corpse in the agent of described worm corpse.
8. the entomopathogenic nematode HbSD suspension preparation described in Claims 2 or 3 for preventing and treating insect, described insect be end age greater wax moth larva, 3 age Sugarcane Yellow snout moth's larva larva, end age Yellow meal worm larva, end age Brontispa longissima larva or end age Clania variegata Snellen larva.
9. the entomopathogenic nematode HbSD worm corpse agent described in claim 6 or 7 for preventing and treating insect, described insect be end age greater wax moth larva, 3 age Sugarcane Yellow snout moth's larva larva, end age Yellow meal worm larva, end age Brontispa longissima larva or end age Clania variegata Snellen larva.
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Cited By (2)
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| CN110199743A (en) * | 2019-07-12 | 2019-09-06 | 浙江绿神天敌生物技术有限公司 | A kind of control method of Spodopterafrugiperda |
| GB2581540A (en) * | 2019-03-25 | 2020-08-26 | Bionema Ltd | Pest control kit and method |
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Cited By (5)
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