CN104059888A - Noctuid granulosis virus, and preparation method and application of preparation containing virus - Google Patents
Noctuid granulosis virus, and preparation method and application of preparation containing virus Download PDFInfo
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Abstract
The invention relates to a virus, and a preparation method and application of a preparation containing the virus, particularly a noctuid granulosis virus, and a preparation method and application of a preparation containing the virus, belonging to the technical field of biology. After the noctuid granulosis virus GXW9-3 is inoculated into larva food of senile noctuid and the like, the larvae can have cytopathy after eating the food; and the carcases collected at 24th-32th hours are disinfected with 75% alcohol on the body surface, pulverized and made into the high-content granulosis virus GXW9-3 preparation. The preparation can be prepared into required concentrations according to the characteristics of occurrence and hazards of the Noctuidae pests on different plants to control the noctuid larvae with great hazards, high overeating property and high chemical resistance. The preparation has the advantages of favorable control effect and long control time, and can effectively lower the noctuid descendant occurrence base number. The preparation has the functions of controlling the outbreak and hazards of the noctuid pests, promoting the ecological equilibrium, protecting the plant growth, enhancing the yield and improving the quality.
Description
Technical field
The present invention relates to a kind of virus, containing preparation method and the application of this viral preparation, relate in particular to mamestra oleracea granulosis virus, containing preparation method and the application of this viral preparation, belong to biological technical field.
Background technology
Since over half a century, with chemical pesticide control insect, agricultural production and income has been made a great contribution.But the problems such as the environment that long-term dependence and a large amount of use chemical pesticides bring and pollution of agricultural products, disruption of ecological balance and food safety also manifest day by day.Reduce to use high malicious persistent pesticide, and promote nuisanceless production or anti-control techniques has just become the task of top priority, it is particularly urgent that the biological control of insect also just seems.Find that approximately more than 200 plant the viral the sixth of the twelve Earthly Branches that parasitizes Agricultural pests, wherein great majority belong to baculovirus, minority is granulosis virus(GV), because insect viruses have single-minded parasitics highly, common a kind of virus only infects a kind of insect, and to he plant insect and people harmless, therefore disturbance ecology balance not, the existing viral pesticide that commercialization is produced on a small scale, majority is used for preventing and treating lepidoptera pest, such as bollworm gypsymoth prodenia litura tent caterpillar small white etc., China the sixth of the twelve Earthly Branches eighties extensive experimentation promote nucleopolyhedrovirus insecticide.The defect of existing biotechnological formulation is that host is narrow, slow, the weak effect of causing a disease, pathogenic agent are subject to illumination effect and the drug effect phase is short, and secondly the above high temperature storage of not resistance to 40 degree of preparation, the short meeting of shelf-lives lose preventive effect.
Summary of the invention
The technical problem to be solved in the present invention is the defect existing for prior art, proposes a kind of mamestra oleracea granulosis virus, and this granulosis virus(GV) can make three kinds or three kinds of pathogenic death of above exigua larvaes, is commonly called as mamestra oleracea granulosis virus.As three-spotted phytometra, oriental tobacco budworm, Plutella xylostella, Dou Autographa spp etc., containing preparation method and the application of this viral preparation, safely and efficiently pest control.
First the present invention provides mamestra oleracea granulosis virus GXW9-3, and on September 20th, 2013, being deposited in its preserving number of Wuhan, China university (Chinese Typical Representative culture collection center) is CCTCCNO:V201340.
Mamestra oleracea granulosis virus Mamestra oleracea granulosis virus GVGXW9-3 is under the jurisdiction of Rhabdoviridae Granulovirus, is a kind of double-stranded DNA Viruses With Inclusion Body.Under JSM-7001F scanning analysis Electronic Speculum, research is found, Viruses With Inclusion Body is the particle of refractivity, rounded, size is 320x189nm, a virus particle is contained in inside, and the DNA that Virion molecules amount is 85.9KB and protein enclosure form, and the infectivity of virus is mainly determined by the integrity of DNA and protein coat, virus particle external parcel the granulin of fine and close lattice, form viral inclusion body.
The present invention is inoculated in mamestra oleracea granulosis virus GXW9-3 in exigua larvae foodstuff, to allow advanced age its predation cause larva cytopathy, and the corpse of falling ill of collecting at 24h～32h is pulverized a granule viral GXW9-3 preparation of initiative high-content after body surface 75% alcohol disinfecting, concrete preparation method is as follows:
The first step, by field and the indoor larva feeding mamestra oleracea granulosis virus GXW9-3 alternately breeding, the larva breeding when field starts to connect mamestra oleracea granulosis virus GXW9-3 while being 3～5 age; 3～5 instar larvae prodenia litura fasting of breeding when indoor industrially 3.5～4.5 hours, again mamestra oleracea granulosis virus GXW9-3 is sneaked in feed, stir, feeding larva, keep 28～38 ℃ of envrionment temperatures, intensity of illumination 2500-3000Lux, relative humidity 70～75%, treats larva cytopathy;
Second step, in cytopathic 24～32 hours of larva, collect there is sharply spasm reaction, metapedes finger tip inserts plant top tissue or leaf margin mesophyll tissue and hangs upside down dead larva corpse, by larva corpse after body surface alcohol disinfecting, until more than 80% corpse body surface, break and flow out after milky dense thick liquid, larva corpse is crushed to particle diameter more than 75nm;
The 3rd step, the larva body fluid of pulverizing out is made to the mamestra oleracea granulosis virus GXW9-3 preparation that is greater than 11,000,000,000 OB/mL.
In the described the first step, select field or the indoor larva breeding.Larva comprises the larva of three-spotted phytometra, oriental tobacco budworm, Plutella xylostella, cabbage caterpillar, prodenia litura, rice borer or mythimna separata, the step of alternately breeding is that indoor industrially is bred the noctuid insects such as prodenia litura after 5 generations, to keep the stable of toxicity evaluation, again the noctuid insects such as prodenia litura are moved on outdoor host crop and bred for 1 generation, the add-on of described mamestra oleracea granulosis virus GXW9-3 is every hundred jin of dry GXW9-3 bacterial strain 100g of feed access.In described step 3, grinding mode is that sand milling is pulverized.
The present invention further provides the application of the preparation that contains mamestra oleracea granulosis virus, described mamestra oleracea granulosis virus GXW9-3 preparation, with adding the agricultural organosilicon of 8mL to make suspension after 500-600 times of liquid of clear water dilution, is sprayed suspension on plant in the dusk of fine day.
Preferred Study On Spraying Condition is 28-38 ℃, intensity of illumination 2500-3000Lux, under the environment of relative humidity 65%～85%, sprays, and prevention effect can reach 100%, and the lasting period reaches more than 60 days.
Because the pathogenic agent of purifying has the characteristic adapting with field environment, there is high temperature resistant high light and pathogenic very by force, nearly ten kinds of current known host insects and tool broad spectrum.The present invention adopts exigua larvae viable cell under the condition of specified temp, illumination and relative humidity, to breed a noctuid granule viral GXW9-3, and utilizes noctuid granule a kind of microbial preparation safe, that contain a noctuid granule viral GXW9-3 efficiently of viral GXW9-3 initiative.Can be according to the feature of noctuidae pests occurrence and harm on different plants, said preparation is mixed with to desired concn, to occurring, harm face is large, gluttony and the strong exigua larvae of chemical resistance are prevented and treated, that advantage of the present invention is that preparation causes a disease is fast, effective, insecticidal spectrum is wide, again because of the GXW9-3 resistance to height of virus and the demonstration of wet high light and better wild nature, there is lasting control effect, prevention and control time long, can effectively reduce noctuid offspring radix occurs.Reach control noctuid insect and break out and endanger, promote the eubiosis, protective plant growth, raising output, the function of improvement quality.
Accompanying drawing explanation
Fig. 1 is the worm corpse of 500 times of liquid control capsicum prodenia lituras of a 11000000000 OB/ml noctuid granule GXW9-3 preparation.
Fig. 2 is that a noctuid granule GXW9-3 is in JSM-7001F scanning analysis Electronic Speculum feature.
Embodiment
Embodiment
The source of mamestra oleracea granulosis virus GXW9-3 of the present invention is that contriver finds the reversal of the natural order of things prodenia litura corpse of falling ill gathering on the wild eggplant of South Mountain, Zhenjiang on August 13rd, 2004 (high temperature high light season).And on September 20th, 2013, to be deposited in its preserving number of Wuhan, China university (Chinese Typical Representative culture collection center) be CCTCC NO:V201340.
The present embodiment is prepared the preparation containing mamestra oleracea granulosis virus GXW9-3 by the following method:
(1), by separation and purification the exigua larvae corpse of falling ill from field, there is a highly pathogenicity noctuid granule viral GXW9-3 through-5 ℃ ,-15 ℃ ,-75 ℃ freeze-dried backs.Because of the difference of freezing front material water content, the freezing time of each temperature is respectively 80-100 minutes, 1700-1800 minute, 1200-1300 minute.
(2) by field and indoorly alternately breed.Be that indoor industrially is bred the noctuid insects such as prodenia litura after 5 generations, to keep the stable of toxicity evaluation, again the noctuid insects such as prodenia litura are moved on outdoor host crop and bred for 1 generation, during the larva in 3～5 ages, start to connect 3～5 instar larvae prodenia litura fasting 3.5～4.5 hours that mamestra oleracea granulosis virus GXW9-3 or indoor industrially breed, again mamestra oleracea granulosis virus GXW9-3 is sneaked in feed, stir, feeding larva, keep 28～38 ℃ of envrionment temperatures, intensity of illumination 2500-3000Lux, relative humidity 70～75%, treats larva cytopathy;
(3) when (2) larva cytopathy, and that at 24h～32h, collects falls ill corpse after body surface 75% alcohol disinfecting, when 80% above corpse body surface breaks the milky dense thick liquid of outflow, with sand mill, be crushed to particle diameter more than 75nm, make a granule viral GXW9-3 preparation of every milliliter of above high-content of GXW9-3 bacterial strain 11,000,000,000 OB, be mainly the larva body fluid that sand milling is pulverized out, sampling coated plate Electronic Speculum is measured the mamestra oleracea granulosis virus GXW9-3 preparation of 110～12,800,000,000 OB/mL.
(4) a granule viral GXW9-3 preparation is carried out to correlation test.
Above larva can be the larva of three-spotted phytometra, oriental tobacco budworm, Plutella xylostella, cabbage caterpillar, Dou Autographa spp, rice borer, mythimna separata.
1 telling test
The morphological specificity of 1.1 mamestra oleracea granulosis virus bodies (MOGV)
A noctuid granule GXW9-3 preparation causes the violent spastic reaction of middle kissing bug generation and climbs to capsicum top branch and leaf margin portion, and the perverse insertion mesophyll of two pairs of foots of rear abdomen is reversal of the natural order of things dead (seeing Fig. 1).
The corpse polypide liquid of falling ill is studied discovery under JSM-7001F scanning analysis Electronic Speculum, virus is the particle of refractivity, circular, size is 320x185nm, 1～2 virus particle is contained in inside, Virion molecules amount is that the DNA of 85.9KB and protein enclosure form, virus particle external parcel the granulin (seeing Fig. 2) of fine and close lattice.
1.2 instrument
Pipettor, graduated cylinder, test tube, slide glass, spirit lamp, WARNING type gold spraying instrument, JSM-7001F scanning electron microscope analysis instrument.
1.3 test method
1.3.1 prepare noctuid particle virus (MOGV) picture
By noctuid particle virus (MOGV) GXW9-3 suspension making, with pipettor, get 1mL, be diluted to 10 ﹣ 5-10-6, then get diluent 0.01mL on 2cm2 slide glass, will on slide glass dislocation spirit lamp, dry fast.
1.3.2 slide spray platinum
By metal spraying in noctuid particle virus (MOGV) GXW9-3 slide sample input WARNING type that carries gold spraying instrument of making, approximately 20 seconds, metal spraying thickness was 20nm, shut down and took out.
1.3.3 the projection of mamestra oleracea granulosis virus particulate image
By being sprayed with the slide glass input JSM-7001F scanning electron microscope analysis instrument of platinum, track down and arrest target, on screen, there is the figure of virion body 320x185nm left and right, be mamestra oleracea granulosis virus (MOGV) GXW9-3 virion.
2 measure containing BO amount and miscellaneous bacteria rate
2.1 treat test sample dilution
Get 8, test tube, in every test tube, fill sterilized water 9mL, with aseptic pipettor draw samples 1mL, add first test tube, aspirate three times, after fully shaking up, pipettor is changed aseptic suction nozzle, draws 1mL and proceeds to second test tube, the like, obtain extension rate and be 101,102,103,104,105,106,107,108 mamestra oleracea granulosis virus diluent.
2.2 slide glass coatings and mamestra oleracea granulosis virus counting
2.2.1 slide glass coating
Get respectively the mamestra oleracea granulosis virus that extension rate is 106,107,108 3 gradients (MOGV) GXW9-3 diluent 0.1mL; be coated on 2*2cm slide glass; repeat 5 times; totally 15; with sterilized water, make blank; to on slide glass dislocation spirit lamp, dry fast; metal spraying in noctuid particle virus (MOGV) GXW9-3 slide sample input WARNING type that carries gold spraying instrument that making is dried; approximately 20 seconds; metal spraying thickness is 20nm, shuts down after taking-up, then will be sprayed with the slide glass input JSM-7001F scanning electron microscope analysis instrument of platinum; track down and arrest target, count.
2.2.2. mamestra oleracea granulosis virus (MOGV) GXW9-3 and miscellaneous bacteria are counted
Under JSM-7001F scanning electron microscope analysis instrument, select the same extent of dilution sheet (5) of mamestra oleracea granulosis virus (MOGV) GXW9-3 number in 30～300 scopes on 2*2cm slide glass, check the mamestra oleracea granulosis virus body sum in 5 sheets choosing.Non-mamestra oleracea granulosis virus body (MOGV) GXW9-3 morphological specificity be counted as miscellaneous bacteria.
Every mamestra oleracea granulosis virus body mean number A=virus sum/5:
Every non-mamestra oleracea granulosis virus body mean number=non-(MOGV) GXW9-3 sum/5.
The mensuration of 3 toxicity evaluations
3.1 materials and reagent
Prodenia litura: catch in field, standby after an indoor feeding generation.
Feed: the not cabbage leaves of pesticide grown or Pepper Leaves.
3.2 measuring method
Employing leaf dipping method is measured.According to the LC50 value of estimating, a upper and lower 2-3 gradient as center, to treat that test sample and standard substance are diluted to 5-7 geometric ratio series concentration, take clear water as contrast, liquid to be measured by the dish leaf of Gong feeding at different concns, in reference liquid He in clear water, soak 5s, after taking out, naturally dry, in every beaker, put into the dish leaf 2-4 sheet after processing, 20 of access third-instar larvaes, in triplicate, 60 of every concentration co-processing larvas, with preservative film, seal, put into 27 ± 1 ℃ of temperature, photoperiod is in the insectary of 14:10h (L:D), 48h checks examination worm death condition, with thin label, touch polypide, nonresponder is dead worm completely.Calculate average mortality and the corrected mortality of testing sample and standard substance.Contrast death is greater than 10%, and test result is invalid, must again test.
Corrected mortality=(chemicals treatment mortality ratio-blank mortality ratio)/(1-blank mortality ratio) * 100% ... (3)
By the numeral input SPSS12 software of test gained, obtain LC50 value and the fiducial limit of standard substance and testing sample, the toxicity evaluation of counting testing sample.
Testing sample toxicity evaluation=(standard substance LC50 value * standard substance are tired)/testing sample LC50 value ... (4)
3.3 tolerance
Toxicity test allows relative deviation, and the maximum relative deviation of three replication results of each sample is no more than 20%.Or it is not remarkable through suitable statistical method checkout discrepancy.
The mensuration of 4 suspensibilities
4.1 instruments, reagent
Graduated cylinder (250mL tool plug), pipette, stopwatch, triangular flask (500mL, 100mL), thermostat water bath, standard hard water (compound method is undertaken by GB/T5451-2001 Plays hard water compound method).
4.2 operation steps are poly-
After sample is shaken up, get 5mL (being accurate to 0.01mL), in 100mL triangular flask, add standard hard water, with hand action circumferential motion, left-right rotary each 30 times.Make suspension and move on in 250mL tool plug graduated cylinder, with standard hard water, be diluted to 250mL.
Graduated cylinder is put into 30 ℃ ± 1 ℃ thermostat water bath, when suspension temperature reaches 30 ℃, graduated cylinder is built, pick up graduated cylinder and shaken gently throw out, the 30s that then turns upside down centered by graduated cylinder middle part regularly, makes to be unit for uniform suspension, graduated cylinder is still put into thermostat water bath, open bottle stopper, standing 30min, extracts the suspension on graduated cylinder top 9/10 out (in 15s～30s, completing) by pipette with degassing method.In drawing liquid process, suction pipe should depend on barrel and decline with liquid level, does not stir bottom precipitation.
With colony counting method measure sample and stay graduated cylinder bottom 25mL suspension containing noctuid particle virus quantity.
4.3 calculate
Specimen suspension rate (%)=111.1 (B-C)/B
In formula:
B-formulated suspension in materialsing containing granulosis virus(GV), unit (hundred million OB);
The bacteria containing amount of C-stay graduated cylinder bottom 25mL suspension, unit (hundred million OB).
(4) .5 pourability test
(4) .5.1 method summary
The suspension agent sample that is placed in container is placed after certain hour, and program is toppled over according to the rules, measures and is trapped in container
The amount of interior sample; After container is washed with water, then measure the sample size in container.
(4) .5.2 instrument
Tool grinding port plug graduated cylinder: 500mL ± 2mL.Graduated cylinder height 39cm, upper and lower scale spacing is from 25cm.
(4) .5.3 testing sequence
The sample of extraction is poured in the graduated cylinder having weighed and (comprised stopper), be filled to 8/10 place of graduated cylinder volume, jam-pack grinding port plug, weighs, place after 24h, open stopper, by graduated cylinder by erect position ROT13 50, topple over 60s, then be inverted 60s, again weighing tank and stopper.
To be equivalent in water (20 ℃) graduated cylinder of falling people of 80% graduated cylinder volume, jam-pack grinding port plug, puts upside down graduated cylinder after 10 times, by aforesaid operations, topples over content, weighs for the third time graduated cylinder and stopper.
(4) .5.4 calculates
Resistates W1 (%) after toppling over, and the resistates W2 (%) after washing, calculate by (6), (7) formula respectively.
In formula:
The quality of m1-graduated cylinder, grinding port plug and sample, g;
After m2-topples over, the quality of graduated cylinder, grinding port plug and resistates, g;
After m3-washing, the quality of graduated cylinder, grinding port plug and resistates, g;
The quality of m0-graduated cylinder, grinding port plug constant weight, g.
6 lasting foaming characteristics tests
6.1 method summaries
The sample of specified amount is mixed with standard hard water, record lather volume after standing.
6.2 reagent
Standard hard water: p (Ca2++Mg2+)=342mg/L, pH=6.0～7.0.Press GB/T14825-2006 preparation.
6.3 instrument
Tool plug graduated cylinder: 250mL (250mL scale marks is to bung's bottom portion 4cm～6cm for scale division value 2mL, 0～250mL scale marks 20cm～21.5cm).
Industrial balance: sensibility reciprocal 0.1g, carrying capacity 500g.
6.4 determination step
Graduated cylinder is added to standard hard water to 180mL scale marks place, put graduated cylinder on balance, be weighed into sample 1.0g (being accurate to 0.1g), add hard water to the scale marks place apart from graduated cylinder plug bottom 9cm, cover plug, centered by graduated cylinder bottom, turn upside down 30 times (each 2s).Be placed on standing 1min on testing table, record lather volume.
7 low-temperature stability tests
7.1 method summaries
Sample keeps 1h at 0 ℃, and observing outward appearance has unchanged.Continuation is stored 7d at 0 ℃, tests the whether conformance with standard requirement of its physical index.
7.2 instrument
Refrigerator: keep 0 ℃ ± 2 ℃.
7.3 testing sequence
Get 80m L sample and be placed in 100mL beaker, in refrigerator, be cooled to 0 ℃ ± 2 ℃, keep 1h, every 15min, stir 1 time therebetween, each 15s, observing outward appearance has unchanged.Beaker is put back to refrigerator, at 0 ℃ ± 2 ℃, continue to place 7d.After 7d, beaker is taken out, return to room temperature, by 4.6,4.7 mensuration that complete suspensibility and wet screening test.Suspensibility and wet screening agreement with experimental standard-required are qualified.
8 heat storage stability tests
8.1 instrument
Thermostat container: (45 ± 2) ℃; Ampoule: 50mL; Injector for medical purpose: 50mL.
8.2 testing sequence
With syringe, by about 30mL sample, inject clean ampoule (avoiding sample contact bottleneck), put this ampoule refrigeration in cryosel is bathed, with thermal-flame sealing (avoiding solvent evaporates).At least seal 6 bottles, weigh respectively.The ampoule of sealing is placed in metal vessel, then metal vessel is put into thermostat container (45 ± 2 ℃), place 42d.Taking-up is wiped away ampoule outside after clean and is weighed respectively, and the sample that quality does not change carries out the mensuration of content and suspensibility in 24h, and after heat storage, effective ingredient rate of decomposition is not more than 5%, and suspensibility conformance with standard requires as qualified.The mamestra oleracea granulosis virus suspension agent package of 11000000000 OB/mL should be stored in ventilation, dry storehouse.
The present embodiment further provides the application of a noctuid granule viral GXW9-3 preparation, mamestra oleracea granulosis virus GXW9-3 preparation, with adding the agricultural organosilicon of 8mL to make suspension after 500-600 times of liquid of clear water dilution, is sprayed suspension on plant in the dusk of fine day.Preferred Study On Spraying Condition is 28-38 ℃, intensity of illumination 2500-3000Lux, under the environment of relative humidity 65-85%, sprays, and prevention effect can reach 100%, and the lasting period reaches more than 60 days.
Preventive effect experiment
(1) the 11000000000 OB/ML granules viral GXW9-3 preparation obtaining is carried out to indoor biometrics test
Toxicity Determination result shows: adopt immersion method to process after prodenia litura primary larva in 3 age, the LC50 value of a noctuid granule viral GXW9-3 sterilant is 0.03110mL/L (95% fiducial limit is 0.0229～0.0423mL/L), shows that Spodoptera litura larvae is had to obvious insecticidal activity; Compare for 0.4422g/L (95% fiducial limit is 0.2836～0.6895g/L) with the LC50 value that contrasts medicament Bt WP (60000IU/mg), its virulence is apparently higher than Bt WP (60000IU/mg), 95% fiducial limit not overlapping (table 1) of LC50 value, statistical study significant difference.
A table 1 noctuid granule viral GXW9-3 sterilant is to the Toxicity Determination result of prodenia litura (leaf dipping method primary larva in 3 age) 2013.8.4
(2) the 11000000000 OB/ML granules viral GXW9-3 preparation obtaining is carried out to the wild snout moth's larva efficiency test of field Chinese littleleaf box leaf roll
A 11000000000 OB/ml granules viral GXW9-3 preparation is for the prevention and control of Chinese littleleaf box leaf roll snout moth's larva, and concentration is advisable with 800 times of liquid left and right, and unit surface dosage is depending on Chinese littleleaf box vegetation or leaf area index, take evenly to spray mist and liquid spreads all over blade as degree.
The control effect of each medicament of table 2 to Chinese littleleaf box leaf roll snout moth's larva
Each medicament of table 2 to the wild snout moth's larva of Chinese littleleaf box leaf roll in natural prevention effect
2013.7.28
Outside each medicament chamber, prevent and treat test-results in Table 1.As can be seen from Table 1,1d after medicine, a 11000000000 OB/ml granules insect population attenuating rate that viral GXW9-3 preparation is processed is the highest, and preventive effect is best, is secondly 48% Le Siben.3d after medicine, the insect population attenuating rate that the short steady bacillus of 10,000,000,000 spores/ml is processed and preventive effect are still according to first, and the preventive effect of 2% first dimension salt, 1.2% kuh-seng nicotine, 48% Le Siben is better.7d after medicine, 48% Le Siben preventive effect is best, reach more than 99%, a 11000000000 OB/ml granules viral GXW9-3 preparation occupy the second, 25% go out No. 3 preventive effects of young urea be extremely significantly worse than other each process, only have 37.26%.From the lasting preventive effect of medicament and the speed that takes effect, a 11000000000 OB/ml granules viral GXW9-3 preparation is best, not only has promptness, has long-lasting simultaneously.
(3) the 11000000000 OB/ML granules viral GXW9-3 preparation obtaining is carried out to field prevention and control test
The GXW9-3 preparation capsicum insect test of table 3,110 hundred million BO/ml granules
2013 month August be prodenia litura on the capsicum in autumn of Zhenjiang, Jiangsu vegetable-growing area just, the season that three-spotted phytometra and oriental tobacco budworm break out, whole capsicum complete stool is injured, three kinds of insect generation overlaps, Sun Jianhua and two senior agronomists of literary composition of higher primary school select a worm state the most complicated, endanger the most serious capsicum field, at 10 o'clock in morning August 16, by 500 times of liquid spray methodes of a 11000000000 OB/ml granules GXW9-3 preparation, prevent and treat, while observing after preventing in 6.30 minutes mornings of August 17, find that the insect that exposes predation all climbs capsicum top branch and leaf margin portion, it is dead that the two pairs of perverse insertion of foot You Nenjitiao face tissues of rear abdomen or mesophyll tissue are reversal of the natural order of things, (a this insect noctuid granule GXW9-3 preparation nocturnal moth class insect climbs capsicum top branch and leaf margin portion, the perverse insertion mesophyll of two pairs of foots of rear abdomen is the reversal of the natural order of things phenomena of mortality, through 6 analysis experts such as the Wu Fuan researcher of Can Ye institute of the Chinese Academy of Agricultural Sciences and medical science professors Lu Rongzhu of Jiangsu University, may be the spastic reaction of middle kissing bug).Because of prodenia litura, three-spotted phytometra and oriental tobacco budworm and have the harm of slipping in pepper fruit and turn green pepper and the feature of the harm of hiding by day and coming out at night, once taking food a pepper plant surface particles body GXW9-3, contact just will certainly die undoubtedly, the community capsicum growing way that adds a sprinkling granule GXW9-3 is light green luxuriant, can lure every day collection to the stronger prodenia litura of the transport property corpse of falling ill, according to manually collecting day by day the record of worm corpse from October 25, (table 3) 17 days～2013 August in 2013
As seen from Table 3, from August 16 with 500 times of liquid of 11,000,000,000 BO/ml granules GXW9-3 preparation control capsicum insect one-factor experiments to the time in 67 days on the 25th October, 500 times of liquid of a 11000000000 OB/ml granules GXW9-3 preparation show as fast, efficient, holding effect and can long-time Control pests and do not injure natural enemy (as schemed).
(4) 110 hundred million OB/ml granule GXW9-3 preparation shop experiment tables of original record
Test period: on May 6th, 2014 was to May 9
For trying material: insect title: small cabbage moth medicament title: a 11000000000 OB/ml granules GXW9-3 preparation
Weaker concn: 1., 2. number sample is respectively the contrast of 600,800,1000 times of liquid clear water, repeat 10 of every processing primary larva in 3 age for three times; 500 times of liquid of a 11000000000 OB/ml granules GXW9-3 preparation, larva in 12 3 ages, 12 primary larva in 4 age
Table 4 result investigation (dead/to live)
Except above-mentioned enforcement, the present invention can also have other embodiments.All employings are equal to the technical scheme of replacement or equivalent transformation formation, all drop on the protection domain of requirement of the present invention.
Claims (8)
1. a mamestra oleracea granulosis virus GXW9-3, its preserving number is CCTCC NO:V201340.
2. a preparation method who contains the preparation of mamestra oleracea granulosis virus, comprises the following steps:
The first step, by field and the indoor larva feeding mamestra oleracea granulosis virus GXW9-3 alternately breeding, the larva breeding when field starts to connect mamestra oleracea granulosis virus GXW9-3 while being 3～5 age; 3～5 instar larvae prodenia litura fasting of breeding when indoor industrially 3.5～4.5 hours, again mamestra oleracea granulosis virus GXW9-3 is sneaked in feed, stir, feeding larva, keep 28～38 ℃ of envrionment temperatures, intensity of illumination 2500-3000Lux, relative humidity 70～75%, treats larva cytopathy;
Second step, in cytopathic 24～32 hours of larva, collect there is sharply spasm reaction, metapedes finger tip inserts plant top tissue or leaf margin mesophyll tissue and hangs upside down dead larva corpse, by larva corpse after body surface alcohol disinfecting, until more than 80% corpse body surface, break and flow out after milky dense thick liquid, larva corpse is crushed to particle diameter more than 75nm;
The 3rd step, the larva body fluid of pulverizing out is made to the mamestra oleracea granulosis virus GXW9-3 preparation that is greater than 11,000,000,000 OB/mL.
3. contain according to claim 2 the method for producing insecticide of mamestra oleracea granulosis virus, it is characterized in that: in the described the first step, larva comprises the larva of three-spotted phytometra, oriental tobacco budworm, Plutella xylostella, cabbage caterpillar, prodenia litura, rice borer or mythimna separata, described alternately breeding is that indoor industrially is bred the noctuid insects such as prodenia litura after 5 generations, then the noctuid insects such as prodenia litura are moved on outdoor host crop and bred for 1 generation.
4. contain according to claim 3 the method for producing insecticide of mamestra oleracea granulosis virus, it is characterized in that: described larva is Spodoptera litura larvae at advanced age.
5. contain according to claim 3 the method for producing insecticide of mamestra oleracea granulosis virus, it is characterized in that: the add-on of described mamestra oleracea granulosis virus GXW9-3 is every hundred jin of dry GXW9-3 bacterial strain 100g of feed access.
6. contain according to claim 2 the method for producing insecticide of mamestra oleracea granulosis virus, it is characterized in that: in described step 3, grinding mode is that sand milling is pulverized.
7. contain according to claim 2 the application of the preparation of mamestra oleracea granulosis virus, it is characterized in that: described mamestra oleracea granulosis virus GXW9-3 preparation, with adding the agricultural organosilicon of 8mL to make suspension after 500-600 times of liquid of clear water dilution, is sprayed suspension on the larval feeding plant such as prodenia litura in the dusk of fine day.
8. contain according to claim 7 the application of the preparation of mamestra oleracea granulosis virus, it is characterized in that: described Study On Spraying Condition is 28～38 ℃, intensity of illumination 2500～3000Lux, under the environment of relative humidity 65%～85%, sprays.
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