TW381003B - Coated pesticidal agents, processes for their preparation and compositions containing them - Google Patents

Abstract

An aqueous process for the preparation of a coated pesticidal agent which process comprises: (a) preparing an aqueous mixture of a pH-dependent polymer and optionally a plasticizer; (b) dissolving the pH-dependent polymer by adjusting the pH of the mixture of step (a) with a base to a pH above the solubilization pH of the pH dependent polymer; (c) adding a pesticidal agent, an ultraviolet protector, optionally a stilbene compound, optionally a disinte-grating agent and optionally a glidant to the solution of step (b), and blending to produce a homogeneous suspension containing dissolved pH-dependent poly-mer; (d) drying the homogeneous suspension of step (c); and optionally; (e) milling the dried material of step (d), wherein said pH dependent polymer is selected from the group consisting of methacrylic acid and methyl methacry-late copolymers, maleic anhydride and styrene copolymers, and mixtures thereof, the pH is adjusted in step (b) to pH 8.5 to 10.

Description

Some pesticides are not activated by the ultraviolet rays of sunlight. Since their insecticides are suitable for controlling unwanted pests, and the areas where they are applied are exposed to ultraviolet rays', a composition containing such agents and being stable to light is needed. In order to prevent the insecticide from being inactivated by ultraviolet rays, the prepared composition contains an ultraviolet absorber and / or a reflecting agent, and an insecticide. U.S. Patent No. 3,541,203 describes a protected virus composition useful for insect control. Preferred compositions include a virus' actinic ray absorbing substance and a polymeric binder substance. This patent states that in order to combine a mixture containing actinic ray absorbing substances and viruses with ethylcellulose polymerizable substances, it is necessary to mix the mixture with ethylcellulose in a toluene solution. The resulting leakage compound is stirred, treated with polybutadiene, poured into a petroleum distillate, and the ethyl cellulose is solidified to produce very small particles of ethyl cellulose polymerizable material, which is substantially uniformly embedded with photochemical A mixture of radiation-absorbing substances and viruses. The particles were then washed several times with another portion of petroleum distillate to completely remove the remaining liquid polybutadiene material. Unfortunately, the method used to prepare the preferred composition of U.S. Patent No. 03,541,203 is not completely printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the notes on the back before filling out (This page)

1T is satisfactory because it requires the use of toxic substances and several steps of washing with flammable solvents. U.S. Patent No. 4,94 8,586 discloses a microencapsulated insecticidal sexually transmitted pathogen. It was pointed out that there are four kinds of micro-embedding compositions that can reduce the light inactivation effect of S. alfalfa (Auto ^ raphe. NPV. However, after exposure to sunlight, the micro-embedding composition retains only the original activity of 30 · 7 to 71.43% β U.S. Patent No. 4,948,586 discloses a method for preparing micro-embedded insecticidal pathogens, which has multiple steps.

A7, B7 11 —-5. Description of the invention (2) Time-consuming and laborious. This method, or microencapsulation of insecticidal pathogens described in U.S. Patent No. 4,948,586, is clearly not completely satisfactory in protecting insecticidal pathogens from the effects of ultraviolet light. One of the objects of the present invention is to provide a coated insecticide that retains a significant amount of its original activity after exposure to ultraviolet rays. The object of the present invention is also to provide a simple and convenient method for preparing a coated insecticide, which is more suitable for commercial manufacture. Another object of the present invention is to provide a wettable powdery insecticidal composition containing a coated insecticide. SUMMARY OF THE INVENTION The present invention illustrates that a coated insecticide retains a significant amount of its original activity after exposure to ultraviolet light. The coated insecticide of the present invention includes an insecticide core, and the periphery is surrounded by a matrix. The matrix contains about 2 to 25 wt% of a polymer that varies depending on pΗ, 0 to about 5 wt% plasticizer, and about 5 to 45% by weight of an ultraviolet shielding agent, 0 to about 75% by weight of a stilbene compound, 0 to about 10% by weight of a disintegrant, and 0 to about 10% by weight of a slip agent. Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) The present invention also provides a method for preparing coated pesticides, and wettable powders containing coated pesticides Insecticidal composition. Brief Description of the Schematic Figure 1 is a diagram illustrating the Ac M NPV genome indicating a specific position of egt. In Figure 1A, the entire A c M N P V genome is represented in map units and is expressed in Eco RI and Hind III restriction maps. Figure 1B illustrates a more detailed map of the region between map units 7.6 and 11.1, and shows the genes Position _ -5 -______ This paper size uses the Chinese National Standard (CNS) M specification (210X297 mm 1 " A7 __B7_ V. Description of the invention (3). Figure 2A is a diagram illustrating the ^ L gene region, in which it is shown In the AcMNpv genome, the key restriction enzyme cleavage positions between the map units 8.3 and 9.8. Figure 2B shows the three forward (1, 2, 3) and three reverse directions of the Ac M NPV genome between the map units 8.3 and 9.8. The structure of the open code frame in the (1, 2, 2, 3,) password (readillg frame). The large open code frame in the code frame 2 indicates the position of the protein coding region of the egt gene. (Carrying) AcMNPV V8 transfer vector NF4 'DNA fragment structure and derivatization diagram. Figure 3A illustrates how fragments a_d combine to form NF4. Figure 3B illustrates the method for preparing fragments c and D. In AcMNPV V 8 £ co RI "I Above the linear limit map of the fragment The arrows indicate the positions of the major open codons (〇RFS) between the map units 032 and 5.83 and the transcription polarity in the AcMNPV genome. The codes “Η” and “Ε” are the restriction endonucleases Hind III and Eco, respectively. Restriction enzyme cutting position of RI. Printed by the Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back before filling out this page). Figure 4 illustrates the structure of pBS ADK-AalT in detail, which contains heterogeneous fat hormones Gene signal sequence and c DNA sequence that encodes AalT and have been adapted by the code and subunit. Figure 5 is a part of the unit expression vector, which contains the expression cassette of the promoter unit, the multi-linker unit and the 3 'UTR unit. Each end contains

Bsu 361 and Sse 83871. The polylinker unit contains an Esp 31 restriction enzyme cleavage site. The area enclosed by the positions of Bsu 361 and Sse 8387 I in the outermost garden is defined as the virus insertion unit β. Figure 6 illustrates the polymerase chain reaction (PCR) method for amplifying fat activity. Chinese National Standard (CNS) A4 regulation; 2HTX297 mm) ~ " " A 7 ___B7 V. Description of the invention (4) Adapt the AalT gene with hormone gene signal / character, and then decompose it with Bam HI. FIG. 7 illustrates that the unit expression vector (AC 0 0 7 5, 1) is formed by adipokinetic hormone gene signal / character adaptation AalT embedded in pMEVl_1, and pMEVl.l contains an AcMNPV DA26 promoter. Detailed description of the present invention The present invention provides a method for preparing a coated insecticide, which can avoid related problems known in the related art. A method of the present invention includes: a) a polymer that varies with pH, an insecticide, a plasticizer, an ultraviolet protection agent as needed, a stilbene compound as needed, a disintegrant as needed, and a slip agent as needed , Blend in an organic solvent such as: acetone, Ci-Cs alcohol, or combine to form a homogeneous suspension; b) take the homogeneous suspension of step (a) to dry; and if necessary c) take the dried substance of step (b) to grind powder. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) The above method of the present invention is beneficial to provide an effective two or three step method for preparing coated pesticides and avoiding the use Multiple mixing and emulsification steps, and avoid several washing steps using flammable solvents. In addition, the method of the present invention described above preferably uses a propylene glycol solution that does not significantly deactivate insecticides such as insecticidal pathogens. Other organic solvents that have been instructed in related technologies such as: Digas methane can significantly deactivate insecticides such as: insecticidal pathogens. Although Xianxin said that the above method has made significant progress over the previous technology, it has been invented. A better way to avoid the use of organic solvents. __— -Ί- This paper size is in accordance with Chinese National Standard (CNS) A4 (210X 297 mm) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7____ 5. Description of the invention (5) It has now been found uniquely. The covered insecticide can be prepared according to an aqueous method, which comprises: a) an aqueous mixture made of a polymer that changes depending on pH and a plasticizer as needed; b) using an alkali adjustment step (a) the mixture to p Η about 8.5 to 10, dissolving the polymer that changes in pH; c) adding an insecticide, a UV protection agent, a stilbene compound if necessary, a disintegrating agent if necessary, and a solution in step (b) 'Use the slip agent and blend to form a homogeneous suspension; d) take the homogeneous suspension of step (c) to dry; and if necessary, e) take the dried material of step (d) and grind it. The advantages of the aqueous method of the present invention are that it does not require the use of multiple mixing and emulsification steps, it does not require the use of flammable solvents for multiple washing steps, and it does not require the use of organic solvents. The particle size of the coated insecticide of the present invention is preferably less than about 20 microns, and the particle size is more preferably about 2 to 10 microns. The homogeneous suspension of the present invention can be dried using any conventional drying technique. Preferably, the suspension is spray-dried or air-dried. The preferred coated insecticides made according to the method described above are those which contain an insecticide core, surrounded by a matrix, and the matrix contains about 2 to 25% by weight of a polymer that varies depending on PΗ, up to about 5% by weight. Plasticizer, about $ to 4 $ weight ° / 〇 UV protection agent 'up to about 75 weight. / Diphenylethylene compound, up to about 10% by weight of a disintegrant and up to about 10% by weight of a sliding agent. The ratio of pesticide to substrate is preferably about 1: 1 to 1: 1. And acetone pair — " —--—_____ * 8 -_____ This paper size applies to Chinese national standards (CNS > Α4 size (210X297 mm))--(Please read the precautions on the back before filling this page), 1T printed by the Consumer Standards Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7, -________ B7_____ V. Description of the Invention (6) The ratio of ci-C 3 alcohol is preferably about 1: 9 to 9: 1 and preferably about 1: 4 to 2: 3 is better. Suitable for CrC with mechanism; alcohols include: methanol 'ethanol, isopropanol and n-propanol, preferably isopropanol. Insecticides suitable for the present invention include chemical properties that will not be activated by ultraviolet rays And biological insecticides, acaricides 'nematicides and fungicides or mixtures thereof. Preferred insecticides are insecticidal pathogens, such as: viral pathogens' bacterial pathogens and fungal pathogens. Applicable virus pathogens include NPV of wild gypsy moth; NPV of Spodoptera litura, such as: AcMNPV E2. AcMNPV L1 ', AcMNPV V8, V8vEGTDEL, and V8vEGTDEL * AaIT; Douglas Fir Tree Nv; European common pine leaf moth NPV; and Heliothis zea NPV. Most suitable for the present invention The insecticides are wild gypsy moth NPV, AcMNPV E2, AcNMPV LI, AcMNPV V8, V8vEGTDEL, V8vEGTDEL-AaIT and corn earworm NPV.

AcMNPV E2 is described in EP 6 2 1 3 3 7 published on October 26, 1994. Polymers suitable for pH change according to the present invention include polymers which are substantially insoluble below about pH 5 such as: methacrylic acid With methyl methyl acrylate, copolymers of maleic anhydride and styrene, and the like, or mixtures thereof. Preferred pH-dependent polymers include copolymers of methacrylic acid and methyl methacrylate, mixtures of methacrylic acid and methyl methacrylate copolymers, and copolymers of maleic anhydride and styrene. The best pH-dependent polymer is Eudragit® S (a copolymer of methacrylic acid and methyl methacrylate, in which the ratio of free carboxyl groups to esters is about 1: 2; Rohm Pharma GmbH, Weiterstadt, Germany) »__- 9 -__ This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) " (Please read the notes on the back before filling in this education) Order A7 B7 5 7. Description of the invention (7) Aiqi L (a copolymer of methacrylic acid and methyl methacrylate, in which the ratio of free carboxyl to ester is about 1: 1, Luomu Pharmaceutical Factory), and mixtures thereof. Plasticizers include any formulation known in the relevant art, such as: polyethylene glycol, polypropylene glycol, diethyl phthalate, dibutyl phthalate, citrate, castor oil, triacetin, etc., or Its mixture is preferably polyethylene glycol with a molecular weight of about 300 to 1000. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling this page) The present invention uses an ultraviolet protective agent to reduce insecticide Light does not activate the agent. Ultraviolet protective agents include ultraviolet absorbers and ultraviolet reflectors, or mixtures thereof. Ultraviolet absorbers include various forms of carbon, such as carbon black, and benzophenones such as 2-hydroxy-4-methoxybenzophenone (CYASORB ®) UV 9, Cytec Ind., 2,2, -dihydroxy-4-methoxyphenone (Hisso UV 24, Hitt), 2-hydroxy-4-propenyloxy Ethoxyphenone (Hisso UV 2098, Hittite) '2-hydroxy-4-n-octyloxyphenone (Hisso UV, 53 1, Hittite), etc .; and dyes such as Congo Red , Malachite green, malachite green hydrochloride 'methyl orange, methyl green, bright green, acridine yellow, FDC green' FDC yellow, FDC red, etc. UV reflectors include titanium dioxide, etc. Better UV protection Agents include carbon black, benzophenone, dyes, and titanium dioxide, with carbon black, Heathrow UV 9 and Heathrow UV 24 being the best. The bases suitable for the aqueous production method of the present invention include: ammonium hydroxide, alkali metal argon oxide, alkaline earth Metal hydroxides, etc., ammonium hydroxide is preferred. Some insecticidal viral pathogens may not be activated at a pH above 10. Therefore It is good to choose a certain amount of alkali and adjust the pH to about 8.5 to 10 to ensure easy dissolution and reduce the chance of inactivation. One of the epiacetyl compounds used in the present invention is to strengthen the insecticidal activity of pesticides. Paper size is applicable to Chinese National Standard (CNS) A4 (210X297). Printed by the Consumers' Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. A7 B7. 5. Description of the invention (8) Properties. Stilbene compounds suitable for this invention are described in US patents. Case No. 5,246,936. A preferred diphenylethylene compound is an analogue of 4,4, -diamino · 2,2, -stilbene dicarboxylic acid, that is, CaCof 丨 〇ur white ( From Sigma Chemical Co., Montana (Sigma Chemical Co., 'St. Louis, MO)' 'e.g. Carcove White M 2R, Carcove White AB, Carcove White LD, Carcove White RWP, etc .; Blancophor (from Mobay Chemical, Pittsburgh, PA)

Chemicals, Pittsburg, Pa ·)), such as: Blankfer BBH, Blankfer MBBH, Blankfer BHC, etc .; INTRAWITE® (a heterocyclic stilbene derivative, From Crompton and Knowles Corp., Charlotte 1 o 11e, NC · Charlotte, North Carolina, such as: Interfacial c F, etc .; Li Ke Leucophor (from Sandoz Chemicals Corp., Charlotte, NC) such as: Li Kefu BS, Li Ke BSB, Li Ke Fu EKB, Li Kefu PAB, Etc .; Pharwite (from Mobike Chemical Company, Pittsburgh, PA), such as: Forbes AR, Forbes BBU, Forbes BKL, Forbes CL, Forbes RKK ', etc. To Blank Forbes BBH Kappa white M 2R and white white AR are the best stilbene compounds. In the present invention, a disintegrant is used to shorten the grinding time and promote a reduction in the particle size of the dry substance. The disintegrants suitable for the present invention include formazan Salts of the condensation products of acidified products with polycyclic aromatic compounds, hydrophilic powders such as dextran, carboxymethyl cellulose, Vinyl pyrrolidine, etc., or its mixture -π-This paper size applies to China National Standard (CNS) A4 (210X297 mm) ----,-1 -------- ΐτ --- --- Lu ·-(Please read the precautions on the back before filling out this page) A7 ---_______ B7____ V. Description of the Invention (9) Compound. The preferred disintegrant is formaldehyde and polycyclic aromatic compound sulfonation. The salt of the condensation product of the product, such as: The salt of the condensation product of formaldehyde and benzenesulfonate, petroleum sulfonate and lignin acid vinegar. (MORWE Ding®) D42 5 (Witco, Houston Texas), Loma (Lomar) pw (Henkei, Cincinnati, Ohio) and Da Where (DARV AN ", (Novak, Conn., RT. Vanderbilt Co., Norwalk, Connecticut) 〇 The coating method of the present invention uses a sliding agent to prevent drying Insecticides stick together. Slippers suitable for use in the present invention include talc, magnesium stearate, calcium stearate, calcium sulfate, etc., or The mixture is preferably talc. The matrix of the present invention may also include other additives such as: preservatives, stabilizers (trehalose), antifungal agents, antibacterial agents, antibacterial agents, etc. ^ Of course, if coated In the case of fungal pathogens and bacterial pathogens, anti-bacteria agents and anti-bacterial agents are usually not used separately. Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the notes on the back before filling this page) The preferred coated insecticides of the present invention are those that contain an insecticide core surrounded by a matrix. The matrix contains About 2 to 20% by weight of a polymer that varies with pH, up to about 3% by weight of a plasticizer, about 5 to 35% by weight of a UV protectant, about 25 to 75% by weight of a phenylethylene compound, and up to about 10% by weight Disintegrant, and up to about 10% by weight of slip agent. The present invention also provides a wettable powder insecticidal composition; comprising about 2 to 25% by weight of a wetting agent; about 2 to 40% by weight of a dispersant; about 100 to 70% by weight of a swelling agent; about 1 Up to 10% by weight flow enhancer; up to about 20% weight __ -12- This paper size applies to China National Standard (CNS) A4 specification (210X 297 mm) ~ ----- Employee Consumer Cooperatives, Central Bureau of Standards, Ministry of Economic Affairs Printed M Β7 V. Description of the invention (10) Amount% p Η modifier; and about 5 to 75 weight? / 0 coated insecticide, which contains the insecticide core, the garden is covered by the matrix, the substrate Then it contains about 2 to 25% by weight of a polymer with a variation of ρ Η, up to about 5% by weight of a plasticizer, about 5 to 45% by weight of a UV-protective agent, up to about 75% by weight of a diphenyl androstene compound, and up to about 10 Disintegrating agent by weight, and up to about 10% by weight of 0/0 sliding agent. Preferred wettable powder insecticidal compositions of the present invention are those which contain about 2 to 15% by weight of a wetting agent; about 2 to 1 5 wt% dispersant; about 10 to 60 wt% bulking agent; about 1 to 5 wt% flow enhancer; up to about 2 Wt% ρ Η modifier; and about 5 to 75% by weight of a coated insecticide, which contains the insecticide core 'is surrounded by a matrix, and the matrix contains about 2 to 25% by weight of the polymer that varies according to ρ Η Substances, up to about 5 wt% plasticizer, about $ 4 to 45 wt.% UV protection agent, up to about 75 wt% diphenylacetamidine compound, up to about 10 wt% disintegrant and up to about 10%. Weight percent slip agent. Humidifying agents suitable for use in the present invention include any formulation known in the relevant art. Preferred wetting agents include anionic agents such as: N-methyl-N-oleyl taurine 'octylphenoxy polyethoxyethanol, nonylphenoxy polyethoxyethanol' 'dioctyl Sodium thiosuccinate, sodium dodecylbenzenesulfonate, sodium lauryl sulfate, sodium sulfonium sulfonate, sodium sulfonate alkyl carboxylate, etc., or a mixture thereof. A mixture of sodium polysulfonate and sodium sulfonated alkyl carboxylate (MORWET® EFW, Witco) is used as the optimal wetting agent. It is suitable for the wettable powder of the present invention. The dispersant of the insect composition includes any conventional preparation known in the related art. The preferred dispersant is an anionic agent such as Kojima scale applicable to China (2ωχ29ϋ (please read the notes on the back before filling out this page) Order A! Printed by the Central Consumers Bureau of the Ministry of Economic Affairs Consumer Cooperatives & Standard (CNS) A4 specification (21 () × 297 Gongqing A7 B7 V. Description of the invention (u): 'Sodium ligninate', which is the condensation product of the acidification product of polymethyl aromatic compounds and polycyclic aromatic compounds, etc. Or its mixture, the sodium sulfonate of tea form condensate is the best, such as: Movitat D425 (Wike Company), Loma Pw (Genk Company) and Davan 1 (R · T. Vanderbilt The company). Bulking agents suitable for the composition of the present invention include natural and synthetic clays and crushed acid salts, such as: natural silica such as diatomaceous earth; magnesium silicate such as talc; Clay and vermiculite; aluminum silicates such as kaolin, montmorillonite, and mica; and hydrated aluminum silicates such as kaolin clay. Preferred expansion agents are hydrated aluminum silicate, broken acid salt, magnesium silicate, and magnesium aluminum silicate. Kaolin clay is the best swelling agent. It is suitable for the wettable powder insecticidal composition of the present invention. The dynamic strengthening agent is a conventional flow strengthening agent known in the related arts, and it is better to use a broken acid salt (such as broken acid). The P Η modifier is used to maintain the aqueous large tank mixture made by the composition of the present invention. Applicable pH modifiers below about pH 5 include potassium hydrogen phthalate and organic acids, preferably citric acid. The method for preparing the wettable powder insecticidal composition of the present invention may be first blended with a wetting agent, dispersant, A mixture of a bulking agent, a flow enhancer and, if necessary, a p (R) modifier, to form a premix. The premix is then blended with the coated insecticide to form the desired wettable powder insecticidal composition of the present invention When used to control insects, the wettable powder insecticidal composition of the present invention is diluted with water to form an aqueous large tank mixture, and the large tank mixture can be directly applied to the pest's nest, food or habitat. The wettable powder composition of the present invention Other ingredients can be added to the product, such as: attractants, gums, defoamers, etc. However, their additional ingredients are usually divided into -14--; ΐτ ^ I (Please read the precautions on the back before filling out this Page) Ministry of Economy Printing Standard A 7 B7 of the Central Standards Bureau Consumer Cooperative V. Description of the invention (丨 2) Add to the large tank mixture. Auxiliaries or adjuvant mixtures can also be added to the large tank mixture. To help further understand the present invention The following examples are mainly proposed in more detail. Except for the scope of patent application, the present invention should not be limited to these examples. Unless otherwise stated, it will be based on "Molecular Colonization: Laboratory by Sambrook et al." Manual, (Molecular Cloning: A Laboratory Manual), 2nd edition, Cold Spring Harbor Press, Cold Spring Harbor, NY (Cold Spring Harbor Press, Cold Spring Harbor, NY) (1989), using standard molecular biotechnology. According to "A Manual of Methods For Baculovirus Vectors and Insect Cell Culture. Procedures" by Summers and Smith, University of Texas 77843-2475, University of Texas Station, Department of Entomology, Texas Agricultural Experimental Station and Texas A & Μ University, (Dept, of Entomology, Texas Agricultural Experimental Station and Texas A & M University, College Station,

Texas 77843-2475)), Texas Agricultural Experiment Station Booklet No. 1 5 5 5 (19 7 8), as a standard technology for the growth and manufacture of virus-like viruses. Example 1 Preparation of Coated V8vEGTDEL Polyhedral Inclusion Body-An Aqueous Preparation Method Add ammonium hydroxide solution (28% NH3) to Agie S 100 (62 g, ROHM) and PEG 400 (6.2 g, polyethylene Diol, average molecular weight 400, Aldrich Chemical Company) in deionized water (551.8 g), until p p is about 9.4. The resulting mixture was stirred for 30 minutes to obtain a solution. Add V 8 ν EGTDEL polyhedral inclusions (62 g, average ___-15 -_____ This paper size applies to China National Standard (CNS) Α4 size (210X 297 mm) (Please read the precautions on the back before filling in this Page) Ordered by the Consumers Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs to print A7 B7___ V. Description of the invention (13) _ ^ The size is about 2 microns, and each gram is about 101 inclusive bodies), Blankferd BBH (248g, Diphenylethylfluorene brightener, MiUs) and Hiso UV 9 (31 grams, average particle size of about 2 microns' Hite Corporation) to p Η adjusted aqueous solution. The resulting mixture was stirred for 30 minutes and air-dried to obtain solid particles. The solid particles were ground by air to obtain coated V8vEGTDEL polyhedral inclusions (362 g, average particle size about 5 microns). The coated VSvEGTDEL polyhedral inclusions prepared above were designated as Composition 1 in Table 11. Basically the same process was used, but the ingredients listed in Table I were used to prepare the coated pesticides of compositions 2-10 in Table 11.

Table I Insecticides a. V8vEGTDEL polyhedral inclusion body

b. Spodoptera litura NPV c _ AcMNPV d. Polymer of 1: 1 mixture of Spodoptera litura NPV and Ac M NPV as a function of pH e. Aiqi S 1 00 plasticizer f. PEG 400 UV-protectant g. Heathrow UV 9 h. Charcoal stilbene compound-16- This paper size applies to Chinese National Standard (CNS) (210X297 mm) " (Please read the precautions on the back before filling this page) Booking 丨 · V. Description of the Invention (Μ) i. Blankfer ΒΒΗ Sliding 剤 j. Talc A7 B7

Table II Coated pesticide composition composition No. pesticide composition prepared by water-based production method / wt / wt% Plasticizer UV-protectant styrenic slipper 1 2 3 4 5 6 7 8 9 10 a / 15.15 a / 24.69 a / 21.74 a / 19.80 a / 17.86 a / 16.53 b / 4.81 c / 21.36 d / 8.41 a / 16.09 polymer e / 15.15 e / 12.35 e / 21.74 e / 9.90 e /17.86 e / 8.26 e / 11.75 e / 9.71 e / 11.98 compound f / 1.52 g / 7.58 i / 60.61 f / l.23 g / 12.35 i / 49.38f / 2.17 g / l〇.87 i / 43.48 f /0.99 g / 9.90 i / 49.41 f / 1-79 g / 8.93 f / 0.83 g / 8.26 f / 1.18 g / 11.75 f / 〇.97 g / 9.71 f / 1.20 g / 11.43 i / 53.57 i / 66.12 i / 70.51 i / 58.25 i / 66.98 (Please read the precautions on the back before filling this page) Order e /3.17 f / 〇.32 h / 32.17 i / 32.17 j / 8.04 -6 丨. Consumer Cooperatives of the Central Standards Bureau, Ministry of Economic Affairs Printing -17- This paper size applies the Chinese National Standard (CNS) A4 specification (210X297 mm) V. Description of the invention (u) Example 2 Production method of coated V8VEGTDEL polyhedral inclusion body-Mechanical method added V8vEGTDEL polyhedron inclusion Body (4324 g average particle size is about 2 micrometers, about 10 1 inclusion body per gram), Blankfer ΒΒΗ (86.44 g) and Hiso UV 9 (31.93 g) to a 30:70 acetone / isopropanol solution (195.15 g) containing IQ 8100 (5.88 g) and PEG 400 (1.51 g). The resulting mixture was allowed to stir for several minutes, then air-dried with stirring to obtain solid particles. The solid particles were ground through a 60 mesh sieve to obtain a coated V8vEGTDEL polyhedral inclusion body, with an average particle size of about 10 microns. The coated V8vEGTDEL polyhedral inclusions prepared above were designated as Composition 11 in Table IV. Basically the same process was used, but the ingredients listed in Table III were used instead to prepare the coated insecticides of compositions 1 2-28 in Table IV.

Table III Insecticides a. V8vEGTDEL polyhedral inclusion bodies b. Polymers of wild gypsy moth NPV as a function of pH c. Iki S 1 00 d. Cypress® 48 (maleic anhydride / styrene copolymerization) (Cytec). Plasticizer e. PEG 400 U V protective agent (Please read the precautions on the back before filling out this page)-Order Printed by the Staff Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs

A7 B7 16 V. Description of the invention (f. Hiso UV 9 g. Charcoal h. T i 〇 2 Diphenyl ethyl hydrazone compound i. Blanker BB Η Disintegrant j. Movit D 4 2 5 stabilizer k. Trehalose

Table IV Composition of the coated insecticide composition prepared by a mechanism method / wt / wt% Composition insecticidal property changes with pH plasticizer UV-protective stilbene disintegrant stabilizer stabilizer No. pathogenic polymer Compound 11 a / 23.76 c / 3.24 e / 0.83 f / 17.55 i / 47.50 j / 7.12-12 a / 70.18 c / 3.51 e / l.75 f / 17.54-j / 7.〇2-13 a / 35.09 c /3.86 e / l; 75 f / 17.54 i / 34.74 j / 7.02-14 a / 23.40 c / 4.91 e / l.75., F / l7_52 i / 45.39 j / 7.02-15 b / 77.19 c / 3.51 e / l.75 f / 17.54---16 b / 62.86 c / 5.71 e / 2.86 f / 28.57--17 b / 40.00 c / 3.64 e / l.82 f / 18.18--k / 36.36 18 b / 77.19 c /3.51. E / l.75 g / 17.54-_ 19 b / 62.86 c / 5.71 e / 2.86 g / 28.57---20 b / 77.19 c / 3.51 e / l.75 h / 17.54----19- This paper size applies Chinese National Standard (CNS) A4 specification (210 X 297 mm) -------__________ 丁 ______ > 0 I _ .- # Λχ (Please read the notes on the back before filling (This page) Printed by the Consumer Standards Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs A 7 B7 V. Description of Invention (17)

Continued Table IV Ingredients of coated insecticide composition prepared by mechanism method / wt / wt% Composition insecticidal property changes with pH Plasticizer UV-protective stilbene disintegrant stabilizer No. Polymerization of pathogens Compound 21 b / 62.86 c / 5.71 e / 2.86 h / 28.57---22 b / 26.83 c / 4.05 e / 2.02 g / 13.42--k / 53.67 23 b / 44.74 c / 2.24 e / l.12 f /ll.22 i / 40.67--24 b / 23.16 c / 2.11 e / l.05 f / 10.53 i / 63.16--25 b / 40.00 c / 3.64 e / l.82 g / 18.18 i / 36.36--. 26 b / 20.37 c / 3.70 e / l.85 h / 18.52 i / 55.56--27 b / 48.89 c / 4.44 e / 2.22 f / 22.22-j / 22.22-28 b / 23.66 c / 3.57 e / l.79 g / ll.83-j / ll.83 k / 47.33 {Please read the precautions on the reverse side before filling out this page) 6n. Applicable to Chinese National Standard (CNS) A4 specification (210X297 mm) A7 B7 V. Description of invention (18) Example 3 Preparation method of wettable powder insecticidal composition Add Table II composition 1 coated V8vEGTDEL polyhedral inclusion body (362g) to containing Movit EFN (13_1g), Movit d425 (26.2g), Kaolin dry soil (91.6g), synthetic Per premix acid (6.5 g, MIC R0_ CEL®E, Manfei Li Company (Manvillo C〇.)) And citric acid (7 g square) of the. The resulting mixture was stirred to obtain a wettable powder composition of composition 29 in Table v. Basically the same process was used to prepare the wettable, powderable composition of τ Composition 30-53 as shown in the table. (Please read the note 4 on the back first to fill in this page), 1T 丨. Printed by the Shellfish Consumer Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs-21 This paper size applies to Chinese National Standards (CNS) Α4 specifications (210 × 297 mm) A7 B7 Description of the invention (19)

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COL srn 000 ss SN 60 εε L9 OS • ετ / 3ΐ a / si 63T 6-6 ε-Β roe / ssεε / sIs Lz 〇inch 6 £ ee LZ 9ε se πεεε εετε 〇ε 6μ 22 This paper is applicable to China Standard (CNS) A4 specification (210X297 mm). Quot " ^ ® ¥ 'o (dwm33a ΓΟυωυμ5.w. ≪ ^ ^ ^ .w. ≪ ® ^ ε (Please read the precautions on the back before filling out this page), 1T A7 B7 V. Description of the invention ( 20)% (NOOOOOO ^ COr- in U) σ ιη σ > 〇〇9 inch · ° 1 inch-SS inch 6 BS 80 89 01 inch one; II ST ST 91 ΔΤ ΔΤ ΔΤ SI step 6 inch · 91 εε · ο 〇 / 〇1 Μ / 1 Μ / φ ^ review ^^^ 絜 ^ 条-畹 ^ Λ ¥ '' M © JMUIOSHS ^^ 辉 ^

sga JgylaMHOW (Please read the notes on the back & fill in this page first) 31 LS 19 95 98 96 £ 6 εε LZ Z inch 88 οε · ε s Δ5 S inch 6L L0 ΙΛ TZ inch ε inch εosos 06 Order

LZ inch ε LT zz 2 9S 93 SN ez SN 61 NT L3 19 ^ s 98 9 ^ ε01 εε 6ε LZ. 3 inch s ετ inch ε ζε ζεε inch? Ε Δε · ετ 59 · ττ ~ Μ! ~ @ HWS ^ OS Δ " 6S LZs inch 8 s LS, 06 08 69 2 18 9 ST IT L OH e IT IT ετετ πτ Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs • οκ-4 ^ · s 9S 66 8Δ T9 QL SS L ^ Ho s • οε / οτ BT / 83 L-LZ VLlZZ zzjzz 91 / 13ετ / s SI / 6I 2/81 zzl L \ es sis 〇s 6 inch 8 inch ίΛ inch 23 This paper standard applies to the Chinese National Standard (CNS) Α4 Specifications (210 × 297 mm) A7 B7 V. Description of the invention (21) Example 4 Evaluation of the insecticidal resistance of irradiated and irradiated wettable powder insecticidal composition against lyre moth (L_dispar) The insect pathogenic composition was suspended in distilled water and diluted to a concentration of 2.4 x 105 coated insecticidal polyhedral inclusions per milliliter. The resulting suspension (05 ml) was pipetted and added to the surface of wheat germ feed contained in a 180 ml plastic cup. Each cup was exposed to ultraviolet light for 0 or 80 minutes (a Westinghouse BLB bulb and a Phillips F40UVB bulb are 3 inches apart, and the distance from the centerline of the light source to the surface of the feed is 4 inches). Larvae are placed in each cup. Cover the cup and keep it dark at 2 9 ° C, 5 5-60 ° RH. After 3 days, the cups were examined to determine the mortality from viral infection. The results are shown in Table Vi, in which the effectiveness of each composition is expressed as the percentage of residual activity (% 〇AR) after the original activity is exposed to ultraviolet light, that is, the mortality% caused by the irradiation < composition + unirradiated Composition-induced mortality x 100. The control composition used in the evaluation is as follows (please read the precautions on the back before filling this page)

1T ^ I. Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 V. Description of the invention (22 Composition composition wt / 1 wt% 1 Coated insecticidal pathogen 11.11 Morwit® ) EF W 13. 5 4 MORWET® D42 5 4., 51 Kaolin Clay 3 0., 12 MICRO-CEL® E 4. 5 1 Sugar 18. 06 MIRA-SPERSE® 18 _ 06 Central Ministry of Economic Affairs Printed by Standard Bureau Shellfish Consumer Cooperative ^ 3.62 wt / wt ° / 〇 Wild Maidu NPV, 4.26% wt / wt ° / o Eudragit® S100, and 2.13 wt / wt% PEG 400. Table VI Confrontation Gypsy moth insecticidal evaluation composition N 0.1 Mortality% Irradiated control group 39 40 41 42 43 44 45 46 47 48 49 50 '51 52 Irradiated unirradiated% OA R2 47.5 100.0 47.5 70.0 100.0 70.0 76.7 99.2 77.2 87.5 99.2 88.2 91.7 98.3 93.2 98-.3 99.2 99.2 57.5) 100.0 57.5 65.0 96.7 67.3 82.5 98.3 84.0 91.Ί 98.3 93.2 90.0 99.2 90.8 90.0 98.3 91.6 8 7.9 100.0 87.9 77.5 99.2 78.2 90.0 99.2 90.7 1 from the combination of Table V Residual Percent Residue of Substance No. 2 after Exposure to UV Scale applicable Chinese National Standard (CNS) A4 size (210X 297 mm) (Please read the back of the precautions to fill out this page)

1T A7 B7 V. Explanation of the invention (23) Example 5 Evaluation of the insecticidal properties of the wettable powder insecticidal composition against HeiC0 Verpa zea This evaluation method uses plastic organisms with 32 open recesses per plate Analytical plate (4x4x2.5 cm, LxWxH, CD International) was used as a test site. 5 ml of Staneville feed (soy / wheat germ) was poured into each recess and allowed to harden. An aqueous suspension (0.4 ml) of a wettable powder insecticidal pathogen composition was hooked on the surface of the hardened material so that each recessed hole contained 4xl05 to 4x107 7 V8vEGTDEL coated or uncoated Polyhedral inclusion body. After the analysis plate was air-dried in a fume hood, a three-day-old cornworm Spodoptera larva was placed on the feed surface of each concave hole. Cover the concave hole with a piece of adhesive ventilated transparent plastic sheet (CD International Corporation), and place it at a constant fluorescent lamp and a temperature of about 27 ° C. After 5 days and 10 days of treatment, the recesses were examined to determine the larval mortality. The results are shown in Table V 11. The control composition used in the evaluation method is as follows. (Please read the precautions on the back before filling out this page) Printed by the Central Consumer Bureau of the Ministry of Economic Affairs Consumer Cooperatives -26- This paper size applies to Chinese National Standard (CNS) A4 (210X297 mm) A7 A7 Central Standard of the Ministry of Economic Affairs Printed by the Consumer Cooperative of the Bureau B7 V. Description of the invention (24) Composition of the control composition wt / wt A Uncoated V8vEGTDEL Polygon 10.00 Body inclusion body Movit MORE WT * EFW 8.53 Movit MOREWT * D425 17.06 Kaolin clay 59.71 MICRO -CEL®E 4,27 citric acid 0.43 B uncoated VSvEGTDEL polyhedral inclusion body 8.69 Movit MORE WT * EFW 8.71 Movit MOREWT®D42f 17.04 Kaolin 60.86 MICRO-CEL®E 4.35 (Please read the back (Please fill in this page again)

* 1T -27- This paper size applies to Chinese National Standard (CNS) A4 specification (210X 297 mm) A7 B7 V. Description of invention (25)

Table VII Evaluation of insecticidal resistance to Spodoptera frugiperdae Mortality ° / 〇 Composition No1 concentration (inclusion body / well) 5 days 10 days Control group A 4x 107 49 91 4xl06 47 93 4x 105 29 63 Control group B 4x 107 33 75 4x 106 35 70 4x 105 18 47 2 9 4xl07 77 100 4x 106 58 100 4x 10s 35 77 3 8 4x 107 43 95 4x 106 45 80 4x 1055 25 56 Composition No. from Table V (Please read the back first (Notes for filling in this page)

* 1T Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs-28- This paper size applies to Chinese National Standards (CNS) A4 (210X297 mm) A7 B7 V. Description of the invention (26) Example 6 Evaluation of the moistened powder composition against Spodoptera litura and H. virescens This evaluation method uses a plastic bioanalytical disc (4x4x2.5 cm, LxWxH, CD International Corporation) with 32 open recesses per disc ) As a test site. Pour 5 ml of Stamfield feed (soy / wheat germ) into each recess and allow it to harden. Take an aqueous suspension (04 ml) of a wettable powder insecticidal pathogen composition and spread it evenly on the surface of the hardened feed, so that each recessed hole contains 4106 pieces of ¥ 8,00 £ 1, and is coated Or uncoated polyhedral inclusions. Take some processed analysis plates under UV light (two Fs 4 OUVB bulbs are placed 30 cm above the analysis plate, Philips Company) or 2 hours. An analysis plate with an estimated irradiation time of 2 hours was selected, and 0.4 mL of deionized water was added to the female recessed holes every one hour to prevent excessive drying and cracking of the copper material. All assay plates were infected with a larva of a three-day large corn earworm or a four-day large American tobacco larvae. Cover the recessed hole with a piece of adhesive-ventilated transparent plastic sheet (CD International Corporation), and place it under a constant fluorescent lamp and a temperature of approx. After 10 days of treatment, the recesses were examined to determine the larval mortality. The results are shown in Table VIII. The control composition used in the evaluation method is as follows. (Please read the precautions on the back before filling out this page) 、 Printed by the 1TB Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs --------- 29-This paper is suitable for financial standards (CNS) A4 specifications A7 B7 V. Description of the invention (27) Control composition

Composition wt / wt ° /. Uncoated V8VEGTDEL Polyhedral inclusion body 4.93 MORWET® EFW 9.0 1 MORWET® D425 18.02 Kaolin dry soil 63.09 MICRO-CEL®E 4.5 1 Citric acid 0.45 Table VIII Unirradiated and irradiated Composition of wettable powder composition against Spodoptera litura and Spodoptera litura No. Irradiation treatment (hours) Evaluation of average larval mortality% Control group 0 Spodoptera litura 59 Spodoptera litura 77 1 46 56 2 40 36 531 0 91 89 1 90 87 2 83 56 (Please read the precautions on the back before filling out this page) Consumption Cooperation by Employees of the Central Bureau of Standards, Ministry of Economic Affairs Du printed 1 Composition No. from Table V Example 7 Solvent Compatibility The sexual assessment method uses the following assessment methods to determine the effects of various solvents and their mixtures on the activity of polyhedral inclusion bodies of Spodoptera litura. A mixture of alfalfa silverworm polyhedra inclusion body (0.55 g) and an appropriate amount of solvent or solvent mixture (5 ml) was taken in a conical test tube or 60 minutes. Centrifuge and decante the test tube ______ -30- This paper size applies Chinese National Standard (CNS) (21〇) < 297 Public Manager A7 --------- B7 V. Description of the invention (28) . The solid was air-dried on a desiccator. Based on the actual irradiation, the process of evaluating the dry solids against the Americas in the night weaving (no results are shown in Table IX. From the data in Table ix, it can be seen that the silver-stained Yee polyhedral inclusions are mixed with digas methane 10 and After 60 minutes, the activity against Spodoptera litura was significantly lower than that of the inclusion body when mixed with acetone, isopropanol, and a 30:70 but isopropanol mixture.

{Please read the notes on the back before filling in this page. J-Order the printed copy of the staff consumer cooperative of the Central Standards Bureau of the Ministry of Economic Affairs. The paper size is applicable to the Chinese National Standard (CNS) Α4 specification (210 × 297 mm) A7 ---_________ ^ V. Description of Invention (29)

Table IX Solvent Propion Solvent Compatibility Evaluation Method Retention Time (Minutes) * Example (Inclusion Body / Concavity) 10 4 x1ο2 Mortality% 19 60 4χ 102 25 10 4 xlO4 100 60 4χ 1〇4 100 Isopropanol 10 4χ ΙΟ2 31 60 4χ ΙΟ2 13 10 4 χ1〇4 100 60 4 χ1〇4 100 Propylene / isopropanol (30:70) 10 4χ ΙΟ2 34 60 4χ IO2 19 10 4 χ1〇4 100 60 4χ 1〇4 100 Digas methane 10 4χ 1〇2 9 60 4χ ΙΟ2 6 10 4 χ1〇4 84 60 4χ 1〇4 19 (Please read the notes on the back first-S to fill in this J)

、 Stamping example of employee consumer cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs 8 Construction of VSvEGTDEL strain To construct a recombinant AcMNPV (V8 strain; ATCC VR-2456) virus that cannot express functional ^ _g_L gene, first contain the AcMNpV gene and cause A DNA fragment extending from the Pst I position of mu7.6 in the AcMNPV genome to the Bam HI position of mu11.1 is embedded in the p st I and Eco RI positions of the p UC 1 9 polylinker in pUC19 (Higma Corporation, St. Louis, Montana). (See Bam _____- 32 -__ of the AcMNPV DNA fragment in Figure 1) This paper size applies to China National Standard (CNS) A4 (210 × 29 · 7 mm) — printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A7 B7 5. Description of the invention (30) Both the HI terminus and the Eco RI terminus of the PUC19 polylinker are treated with T4 DNA polymerase and then filled in, and then the fragments are ligated to promote fragment joining. In this way, the Eco RI position in the polylinker region of the product is destroyed. The resulting plastids are called pUCBCPsB. The pUCBCPsB was cleaved by restriction endonuclease Xba I (see the arrangement of these positions in the §_ ^ gene in Figure 2), treated with T4 DNA polymerase, and the protruding end was connected to X ba cleavage site. Then the linearized plastids were cleaved with Eco RI, and the small Xba I / E c RI fragment was discarded. 'Then E. coli Lac_z was excised from pSKS 104 (3) with Eco RI and Ah a III. The gene 'is embedded between Eco RI and the inserted Xba I position. The resulting plastid is called pEGTZ. In this plastid, the embedded lacZ gene and the leading egt coding sequence are in the code frame ° or, simply Connect Eco RI and Xba I positions together (only after these two positions have blunt ends) 'You can construct the pEGTDEL without any sequence in between. Then as described by Shanmus and Smith (2) The pEGTz and AcMNPV V8 DNA are co-transfected into SF cells. This process can cause homogeneous recombination between the virus and the plastid DNA sequence, resulting in the replacement of the e gj,: J ac Z gene from pEGTZ. Virus egt. Because the remaining coding sequences and L§ ^ Z sequences are in the code frame, the virus produced (called V8vEGTZ) can produce fusion proteins, and its performance can be shown in the chromogenic indicator (eg: 5 -Bromo-4 -Ga-3-Θ 丨 Indolyl-dot · D · Galanose Blue virus plaques are produced in the presence of (X "gal)). Recombinant virus V8vEGTDEL is produced by infecting sf cells with pEG and DEL and DNA from virus VSvEGTZ. Homogeneity __-33-This paper size applies to China National Standard (CNS) A4 (210X 297 mm) '-------- (Please read the precautions on the back before filling this page}

Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs, 1T A7 ____B7 V. Description of the invention (31) After the reorganization, the ^ fusion gene in V8vEGTZ was changed to the missing gene replacement from pEGTDEL. Recombinant virus V8vEgtdel cannot form blue spots in the presence of X · g a 1 and can be judged from this point. Example 9 T-NF002, which is an occ · / ^ _gap derivative of V8vEGTDEL, was constructed before constructing a recombinant derivative of V8vEGTDEL-AalT, first constructing an intermediate virus vector, in which the polyhedron gene has been partially deleted and has been partially removed by the large intestine Bacillus ί_9 ^ _Z gene replacement. The obtained virus could not form a virus fusion (fusion-negative, 0 ° C, and a blue virus spot appeared in the presence of a color-producing indicator X_gal (万 _ga 丨 +). T-NF002 was constructed by V8vEGTDEL dNA and transferred The vector PVL9 41-500 cold gai was co-transfected into SF cells, and some of the AcMNPV polyhedron genes have been replaced with coliform genes (4). When transfected, Cheng Ye was spread on SF cells containing a chromogenic indicator. At the time, there was a blue spot without virus fusion, which was Ding_NF002. Example 10 Construction of an unborne AcMNPV V8 transfer vector NF4 The design of the transfer vector NF4 was based on the use of homogeneous DNA recombination at a position of 92 bp upstream of the polyhedron gene. Function to promote the embedding of foreign genes into the AcMNPV V8 genome. When constructing ^ 4, an 8 bp Bgl π linker (5 '-C AG AT CTG-3' was used; Braunmanham, Indianapolis, Indiana Company (B〇ehringer. Mannheim, indianQpQli% TN)) embedded in the pBluescript® II KS (Stratagene, La JolU, CA)) unique Ec〇 _____ 34 _ this k scales apply; national standards (& q uot; cNsj grid (210X 297 public capital) '~~ --- (Please read the precautions on the back before filling this page) Order A7 Β7 V. Description of the invention (32) The RV position, thus destroying the Eco RV position. This plastid is called AC 0039.1. Then, an Eco RI fragment (Eco RI fragment "I" in Fig. I) extending from 0.32 mu to 5.83 mu in the AcMNPV V8 genome was selected and cloned into the unique Eco RI position of AC0039.1 to form Intermediate plastid NF3. NF 4 is derived from NF 3, which is formed by a 2 2 bp double-stranded DNA sequence ("B su -Sse linker") embedded in NF3 at a position 92 bp upstream of the AcMNPV V8 polyhedron gene. Embedding The following sequence contains the recognition positions of restriction endonucleases Bsu 361 and Sse 83871, as shown below:

Bsu36I Sse8387I 5,-CCTCAGGGCAGCTGCCTGrAdd-3 r 3 '-GGAGTCCCGTCGACGGACGTCC-5, (SEQ ID NO: 1) Since AcMNPV V8 does not contain a suitable restriction enzyme cleavage site at the desired Bsu-Sse linker insertion point, it is shown in Figure 3A The four DNA fragments indicated by AD in the same time are connected at the same time: make up NF4. All fragments were purified by agarose gel electrophoresis. Printed by Shellfish Consumer Cooperation of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the notes on the back before filling this page) The preparation of fragment A is to limit the endonuclease B pu 1 1 0 2 I to decompose NF 3 and to calf intestine Alkaline phosphatase dephosphorylates its ends and cleaves the DNA with Xba I to isolate a large (6.1 kb) Xba I / BpuU02 I fragment. Fragment B was prepared by decomposing N F 3 with Hind 111, dephosphorylating the end with calf intestinal alkaline phosphatase, and cleaving the DNA with Xba I to isolate a 3.26 kb Hind III / Xba I fragment. The manufacturing method of fragments C and D shown in FIG. 3B uses A c M NPVV 8 E c 〇R1 "I" fragment in NF 3 as -35- This paper size applies Chinese National Standard (CNS) A4 specifications (210 × 297 mm) A7 B7 _ 5. Description of the invention (33) DNA template, DNA fragments are synthesized by PCR amplification. The primers used to synthesize fragment C are: V8-1: 51-GCTCTGACGCATTTCTACAACCACGACTCC-3, (SEQ ID NO: 2)

Sse8387I Bsu36I V8-2: 5'-TAATcctqcaqqcaacbqccctqagqATCAGCAACTATATATTAA-GCCG-3 '(SEQ ID NO: 2), printed by the Employees' Cooperative of the Central Bureau of Standards of the Ministry of Economic Affairs (please read the precautions on the back before filling in this page) Introduction V8- 1 is the forward primer in the PCR reaction, which will hybridize with the complementary DNA strand of AcMNPV V8 genome located at mu 2.65. Primer V8-2 consists of two parts. Residues 1 to 4 and 27 to 49 (indicated in upper case in the figure above) are linearized with complementary AcMNPV V8 DNA sequences located 89 to 92 bp and 93 to 115 bp upstream of the translation start position of the polyhedron gene, respectively. This primer segment is indicated by the black frame in the primer V 8-2 of Fig. 3B. Residues 5 to 26 of primer V8-2 (indicated by the underline in the figure above), including the complementary strand of the Bsu-Sse linker sequence, will be embedded 92 bp upstream of the translation start position of the polyhedron gene in the AcMNPV V8 genome. This primer segment is not complementary to any native AcMNPV DNA sequence, and is indicated by the white box in primer V8-2 of Figure 3B. Following the PCR synthesis method, fragments were decomposed with Hind III and Sse 83871 to generate fragment C, which was purified by gel electrophoresis. The primers used to synthesize fragment D are:

Sse8387I V8-3: 5 '-getgcctgcaggATTATGTAAATAATTAAAATGATAACCAT-CTCGC-3' (SEQ ID NO: 4) PVLReverse: 5 '-GGATTTCCTTGAAGAGAGAGTGAG-3' (SEQ ID NO: 5) ____- 36 -__ This paper standard applies to Chinese national standards ( CNS) A4 specification (210X297 mm) A7 B7 Paper size of printed form for employee consumer cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs, China National Standard (CNS) M specification (210 > < 297 mm) V. Description of the invention (34 primers) V 8-3 is composed of two parts. Residues 1 to 12 (indicated by the underline in the figure above) include the 3 'half of the front strand of the Bsu-Sse linker and will not be complementary to any natural A c Μ NPVDNA sequence This primer segment contains only the cleavage position of restriction enzyme 83871 and is indicated by the white box in primer V8-3 of Figure 3B. Residues 13 to 46 are located upstream of the start position of the polyhedron gene translation The 92-59bp AcMNPV V8 DNA sequence is co-linearized. This primer segment is represented by the black box in primer V8-3 of Figure 3B. The primer PVL Reverse is located at the start position of the polyhedron gene translation '' downstream 205 AcMNPV V8 genome complementary strand to 226 bp After the PCR synthesis method, the fragments were decomposed with Bpun〇2 I and Sse8387 I to generate fragment D, which was purified by gel electrophoresis. Equal molar fragments AD (25 femtomoles each) were used at 15. (: 10 μl containing 20 mM Tris-HCl (pH 7.5), 10 m Μ M g C 12 '1 mM DTT, 0-5 mM ATP, and 20 units of T4 DNA ligase (Beverly, Md. Cultured in the ligation solution of New England Biolabs (New England Biolabs, Beverly, MA) for 16 hours to synthesize NF4. After the contents of the ligation reaction were transformed into E. coli, the restriction enzyme analysis method was used to identify the NF4 structure. Example 1 1 The construction of a recombinant virus insecticide containing a gene cassette encoding the cDn A sequence encoding AaIT containing fat from the tobacco hawk moth (M_a nduca s.exta) fat using hormone gene signal sequence and adapting it to a virus, Especially a morbid virus, in which one or more kinds of insect-specific toxins (5_7) and peptide hormone -37- are added (please read the precautions on the back before filling this page). A7 B7 Five The invention description (35) (8) or enzyme (9) foreign genes to enhance its insecticidal properties. The peptide AalT found in North African brown (An dr ct on_us. Venom) is one of the examples of this insect-specific toxin (6). When AaIT is injected into the body cavity of insect larvae, it will interact with voltage-sensitive sodium channels. Selective binding, 收缩 causes contractile paralysis. Insect larvae infection will produce A a 丨 T virus for chronic toxin administration, which can prolong the state of mochi and eventually lead to death. Synthesis of a gene cassette encoding mature AalT toxin, It is connected to the insect signal peptide from the tobacco fat moth's fat-applying hormone (ADK) gene, and is composed of two parts: "B 2" fragment, which is composed of DNA encoding a DK signal sequence plus amine in the toxin coding region. The basal portion, and "A ,, fragments, are the remainder of the coding region encoding toxins. Each fragment A and B is a pair of oligomers containing 15 bp overlap (purchased from Beverly, Maryland) New England Biological Laboratory, New York City) Adhesion formation. SequenaseTM 2_0 (a type of DNA polymerase) (State Biochemical Corporation, Cleveland, OH) Strand molecule, which contains the ADK signal sequence plus a cDNA sequence that is adapted by a coder and encodes AaIT. The list of oligomers used in constructing this construct is as follows: -38- This paper size applies to Chinese National Standard (CNS) A4 Specifications (210x297 mm) (Please read the notes on the back before filling this page)

Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs A7 B7 V. Description of Invention (36)

Fragment A

El 5 'AGCCCCCGAG TGCCTGCTCT CGAACTATTG

CAACAATGAA TGCACCAAGG TGCACTACGC

TGACAAGGGC TACTGTTGCC TTCTGTCCTG CTATTGCTTC 3 '(SEQ ID NO: 6)

E2 5 'CTGTAGGTAC CGGATCCTTA GTTAATGATG

GTGGTGTCAC AGTAGCTCTT GCGAGTATCA

GAGATTTCCA GAACTTTCTT GTCGTCGTTG AGACCGAAGC AATAGCAGGA 3 '(SEQ ID NO: 7) Fragment B2

Common 5 'AGGCACTCGG GGGCTTTTCC GGATGAGGTC GACTGCGTAG CCGTTCTTCT T 31 (SEQ ID NO: 8)

ADK 5 'CCCCCCGGAT CCATGTACAA ACTGACCGTC TTCCTGATGT TCATCGCCTT CGTGATTATC GCTGAGGCCA AGAAGAACGG CTAC 3' (SEQ ID NO: 9) Figure 4 illustrates a method for constructing a gene cassette containing the AalT gene and the tobacco hawk moth ADK signal sequence. In the first step, the "Common" oligonucleotide and "ADK" were bonded, and SequenaseTM 2.0 was used to fill the single-stranded area. The filled-in fragment consists of 12 nucleotides upstream of the non-coding region, which contains the Bam HI position required for breeding, the coding region of the ADK signal sequence, and the first 15 amino acid terminal amino acids encoding AalT. Nucleic acid. This small segment of double-stranded DN A was decomposed by Bam HI and Ava I, and was sub-selected to pBluescript S K + (Starkin Company, La Jolla, California) that had been decomposed by Xma I and Bam HI. Pvu 11 fragments larger than 4 4 5 b p fragments were found only in pBluescript S K +, and therefore positive subpure lines were judged to be present. There is a pair of base pairs between the protrusions at the positions of Ava I and Xma I ___- 39j_ This paper size applies Chinese National Standard (CNS) A4 specification (210X 297 mm) n ϋ-IT m--n _ _ mn _ _ _ T,.-(Please read the notes on the back before filling out this page) A7 V. Description of Invention (37) The consumption cooperation of employees of the Central Procurement Bureau of the Ministry of Economic Affairs does not match printing. Pure strains that have corrected this mismatch are selected by appropriate restriction enzyme decomposition. In Figure 4, the resulting plastid is called pBS ADK B2. When constructing the A fragment encoding the large AalT coding region, the oligos are spliced: y: acids "E1" and "E2", and filled into the single-stranded region using SequenaseTM 2.0. The result was a double-stranded molecule with an 8 "position at the 5, end, followed by the 13am HI / & Asp 718 position after the 3 'end of the toxin coding region. The Bam HI and Asp 718 positions were introduced to assist in turtle selection. Fragment A was decomposed with Ava I and Asp 718. The plastid containing B2 fragment, pBS ADK B2, was also decomposed by Ava I and Asp 71 8 and the fragment a was selected and cloned into the decomposed plastid. In Figure 4, the obtained The plastids are called pBs ADK · AalT. After fragment A is ligated into pBS ADK B2, the ligated DNA is used to transform competent DH5a coli. The small preparation of plastid DNA is prepared from the bacterial colonies. Restriction enzyme analysis was used to determine the colonies containing the desired recombinant DNA. Restriction enzyme analysis was followed by DNA sequence analysis to confirm the completeness of the plastid pbs ADK-B2. Example 1 2 ADK-AalT gene card The ADK.AaIT gene cassette of Example I! Was isolated from pBS ADK-AalT in the occ-defense virus transfer vector as a Bam HI fragment for colonization to alkaline phosphatase that has been degraded and passed through calf intestine. Treated p VL9 8 5 baculovirus transfer vector DNA (4). The resulting plastid is called pVL9 8 5 / ADK-AaIT. Restriction enzyme analysis is used, and then the sequence of the insert is analyzed to determine the correct orientation and integration of 985 / ADK-AaIT. • The 40-sheet scale applies Chinese national standards ( CNS) M size (210X297mm) (Please read the precautions on the back before filling this page)

1T * tn _ Printed by the Consumer Cooperatives of the Central Bureau of Standards and Mobilization of the Ministry of Economic Affairs A7 —__ B7 V. Description of the invention (38) A sample of E. coli strain HB101 containing the transfer vector AC 0055.1 has been deposited with the recipient in the United States by this application Culture Collection (American Type Culture Collection, 12301 Parklawn Drive, Rockville, Maryland 20852, USA) and designated as ATCC number 69166. AC 00 55.1 contains a gene cassette, in which the a a I T gene sequence of Example 11 is connected to the epidermal gene signal sequence of Drosophila, but not to the hormone gene signal sequence of tobacco moth fat. Relevant art experts can use this storage material to use the above-mentioned fat gene hormone gene signal sequence. Replacement of the epidermal gene signal sequence, its DNA sequence is as follows: Epidermal 5 'CCCCCCGGAT CCATGTTCAA GTTCGTGATG ATCTGCGCCG TCCTCGGCCT GGCTGTGGCC AAGAAGAACG GCTAC 3' (SEQ ID NO: 10) Example 1 3 Embed the ADK-AalT gene cassette into the baculovirus unit expression vector Since the ADK-AalT gene cassette in the pVL985 / ADK-AaIT transfer vector of Example 13 replaces part of the virus polyhedron gene, the use of this transfer vector is limited to the construction of a fusion negative (occ_) recombinant virus that can express AalT . It is also suitable for embedding ADK-Aal D in the AcMNPV genome so that the recombinant virus can form a virus fusion. One of the positions suitable for this purpose is the position of the Bsu-Sse connector in the (unsupported) AeMNPV V8 transfer carrier NF4 of Example 10. This position is between the virus polyhedron and the ORF 603 open code box and does not interfere with any known viral functions (see circle 3). However, because the ADK-AalT ^ of Example 12 does not contain the ADK-AalT mRNA's greening effect 3, the treatment (polygland __ -41 -__ This paper scale is applicable to the Chinese National Standard (CNS) A4 specification (210χ_297mm) ()) --- (Please read the notes on the back before filling this page), tr A7 B7 V. Description of the invention (39) Answer the acidification) necessary regulatory sequences, so you must first insert the adk_ AUT gene card into The expression vectors for these important tritium elements can be provided. One such vector is pMEVi.i, which is a unit expression vector based on the general design shown in Figure 5 with Esp 31 as the main unit. pMEV1 ^ is constructed in PB 丨 uescriPt KS- and includes the following components: (1) virus insertion unit, which is delineated by the restriction positions SSe 8 3 8? 1 and Bsu36! The promoter unit, if it is pMEV1.1, the unit is composed of the 5 'untranslated region of the promoter * AcMnpv gene (10); (3) a polylinker unit, which is used to promote genes that can be expressed as required The cassette is inserted; and (4) 3, UTR (Untranslated Region), which provides a place for processing of 3, and polyadenylation of mRNA. pMEVl.I-3, UTR is composed of the AcMNPV basic protein (6.9K) gene, 3, UTR (u). A sample of E. coli strain DH5 a (AC0064.1) containing pMEVl.l has been deposited by the assignee of this application at the American Type Culture Collection (Americal Type Culture C () on April 7, 1993 ilecti (? n, 1 2 3 0 1 Parklawon Drive, Rockville, Maryland 20852, USA), and the instruction number is ATCC 69275. When the ADK-AalT gene card is inserted into pMEVl.l, first use PCR from pVL985 of Example 12 The ADK-AalT gene cassette was recovered from the / ADK-AaIT transfer vector. The pcR method is shown in Figure 6. The (+) strand primer used in this reaction was the oligonucleotide PD30, the 5, end of which was associated with the ATG of the ADK-AalT gene The translation start code is the same. The sequence of pD30 is as follows: PD30 5 'ATGTACAAACTGACCGTCTTCCTGATG-3, ---- 42- This paper size is applicable to China National Standard (CNS) A4 specifications (210X297 public holidays (please read the precautions on the back first) (Fill in this page)

* 1T Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs and printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs and printed by A7 ___B7__ V. Description of the Invention (4〇) (SEQIDNO: ll) The primers for each (-) share are pVL Reverse (SEQ (ID No: 5) (see Example 10), can hybridize with a position of 205 to 226 bp downstream of the translation start position of the polyhedron gene, or approximately 35 bases downstream of the insertion position of the ADK-AalT gene in the pVL985 / ADK-AaIT template Base hybridization. For the amplification reaction, each primer used 50 picograss ears and 250 picograms of pVL985 / ADK-AaIT template DNA, and contained 10 mM Tris-HCl (pH 8.3), 50 mM KC1, 1.5 mM MgCl2, 200 '" 111 <; 1 > 1 Ding? 8. 100 μg / ml gelatin and 2.5 units of AmpliTagTM DNA polymerase (Perkin-Elmer Cetus, Norwalk, CT) in 50 μl of reaction mixture mixing. The reaction was carried out at 25 ° C for 1 minute (variation step), 55 ° C for 1.5 minutes (adhesion step), and 72 ° C for 3.0 minutes (elongation step). It was then extended at 7 2 ° C for 7 minutes. EDTA was then added to 10 mM and Sarkosyl (N-laurate sarcosinate) to 0.2% (w / v) to stop the reaction. The product was extracted once with phenol: aerosol and precipitated with ethanol. After the products were collected, in the presence of all four dNPTs, they were treated with Klein fragments of E. coli DNA polymerase to ensure that the PCR products had 5, blunt ends.

Based on the decomposition products of B a m 定 I, the 3 and ends of the ADK-AalT toxin gene cassette were identified, and the ratio was 18. On agarose gels, electrophoresis was performed to purify the fragments containing ao-κ-AaIT. When preparing p Me V ι · ι for toxin insertion, the DNA is decomposed with β§ρ, and in the presence of all four dNTPsts, the large intestine rod is used. -----43- This paper size is applicable China National Standard (CNS) A4 specification (210X 297 public holiday one) --- ----- (Please read the notes on the back before filling this page),-° A7 B7 V. Description of the invention (41) Bacteria DNA polymerase 1 (Klen fragment), fill in the resulting 5, protruding ends. Then, Na A was decomposed with B a m Η I and electrophoresed on a 1% low-melting agarose gel to separate the carrier from the released multi-linker fragment. The blunt / Bam HI vector fragment was ligated with the equimolar blunt / Barn HI ADK-AalT fragment and transfected into E. coli. The resulting plastid was called pMEV1.1 / ADK-AaIT-1 (AC00 7 5.1), and contained the ADK-AalT coding region controlled by the promoter of the AcMNPV DA26 gene. A diagram of the virus insertion unit in pMEVl.l / ADK-AaIT-l (AC 0075.1) is shown in FIG. Example 1 4 Construction of recombinant virus V8vEGTDEL-AaIT Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs (please read the precautions on the back before filling in this tile) VSvEGTDEL containing ADK-AalT gene controlled by the AcMNPV DA26 promoter In the case of the recombinant derivative, the restriction enzymes bsu 3 6 j and 88683871 were used to decompose plastids 1 > 乂 1.1 / 80 01 ^ 8 & 1 but-1, and DA26 / ADK- containing DA26 / ADK- was purified by agarose gel electrophoresis. AaIT performance card g virus to insert units. This fragment was embedded in the Bs.u_Sse linker of the transfer vector NF4 of Example 10. The resulting transfer vector is called NF5, and contains 9 2 上游 upstream of the start position of the polyhedron gene translocation.) 八 26 / 八 01 ^-八 & 1ding performance card. Therefore, the transcriptional polarity of the AalT gene Contrary to the green polarity of the polyhedron gene. When constructing a recombinant virus containing the AalT gene, the process (2) described by Summers and Smith was used to co-transfect SF_9 virus DNA and the transfer vector NF5 into SF9 cells. Because the T-NF002 virus of Example 9 contains the E. coli gene instead of virus polyhedron-44-This paper is in accordance with the Chinese National Standard (CNS) to the specifications (210 X297 mm) Printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs A 7 · _________B7_ V. Explanation of the invention (42) Because of this, when the virus grows on Sf9 cells in the presence of a chromogenic receptor X_gal>, it is discriminated by a colorless fusion-positive plaque such as w 0 呀 氏 切 ^ 木Recombinant between TNF 0 02 and transfer vector NF5. This virus is called and contains DA26 / ADK-AaIT, which is located in the 92 leaf upstream of the fully functional polyhedron gene. Example 1 5 Preparation method of V8vEGTDEL-AalT fusion. Sf9 cells were infected in the culture, and the fusion was collected 5 days after infection. That is, a fusion (polyhedron) was formed from V8vEGTDEL. In short, the sf9 cells were maintained in complete TNM-FH medium (prepared by the method of Ethan Moss and Smith (2)), in terms of 0.3 x 106 to 4 x 0 per milliliter. Density, maintain log phase growth in rotating culture at 27 ° C. When the fusion was produced, the culture was repeatedly infected with V1vEGTDEL_AalT budding virus at a density of 1.0 × 6 cells / ml in 01_1 () plaque forming units per cell. At 27. (: After 5 days of incubation, in a Beckman GPR centrifuge at 2000 rpm for 10 minutes at 2000 rpm, collect intact cells, cell debris and fusions. The pellet is resuspended in 5 〇

Tris-HCl (pH 7.5), 10 mM EDTA, 0.1% (v / v) Triton X-100, equivalent to 107 cells / ml. Sodium lauryl sulfate (s D s) is added to a final concentration of 1% (w / v), and the released cells d N A are cut and sheared by intense suction. Centrifuge at 2 000 rpm for 10 minutes and collect the fusions at 5 (3 mM

Tris-HCl (ρΗ7.5), 10 mM EDTA, 0.1% τ rit ο η X-1 0 0 Centrifuge for washing twice and then resuspend in sterile distilled water with a density of about 1 ml per fusion. The amount is mainly in the range of 2xl01Q fusions per liter of original culture. _____-45-"This paper is suitable for Zhongguanjiatizhan (CNS) A4 size (2! 〇Χ 297mm ~~~~ (Please read first (Notes on the back then fill out this page)

A7 B7 V. Description of the invention (43) References within the scope Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs 1. Sambrook, J., et al., Molecular Cloning: A Labora Dvororv Manual, 2nd ed., Cold Spring Harbor Press, Cold Spring Harbor, NY, (1989) 2. Summers, MD, and Smith, GE, A Manual Of Methods For Baculovirus Vectors and Insect. Cell CulLure Procedures, Dept. of Entomology, Texas Agricultural Experimental Station and Texas A & M University, College Station, Texas 77843-2475, Texas * Agricultural Experiment Station Bulletin No. 1555 (1987). 3. Casadaban, MJ, et al., Methods Enzvmol., 100, 293-303 (1983). 4. Luckow , VE, and Summers, MD, Virology. 170, 31-39 (1989). 5. Tomalski, MD, and Miller, LK, Nature, 352, 82-G5 (1991). 6. Zlotkin, E., et al ., Toxicon. 9, 1-8 (1971). Ί. Stewart, LMD f et: 'al., Nature, 352. 85-88 (1991).', 8. Maeda, S., B 丄 ochetn. Bit > phvs. Res. Commun. 16 5, 1177-1183 (19Θ9). 9. Hammock, BD, et al., Nature, 344, 458-46 1 (1990). 10. O'Reilly, DR, et al., J. Gen. Virology, 71, 1029-1037 (1990). 11. Wilson, M. E., et al., J. Virology, 61 , 661-666 (1991). -46- The paper size is applicable to the Chinese National Standard (CNS) A4 specification (21 × 297 mm) (please read the precautions on the back before filling this page) Central Standards Bureau of the Ministry of Economic Affairs Printed by employee consumer cooperative A7 Β7 V. Description of invention (44) Sequence list (1) —general information ① Applicant: Archimedes' Fakruddin 7 (Alimed, Fakhruddin) (ii) Name of invention: Coated Insecticides, methods for their preparation, and compositions containing these insecticides. Qiu) Number of sequences: 11 (iv) Correspondence address (A) Address: American cyanamide company (B) Street name: One Cyanamid Plaza (0 City name: Wayne) Chenzhou Name: Group Jersey (N ew Jersey) (E) Country Name: United States (F) Postal Code: 07470 (v) Computer Readable Form (A) Media Type: Floppy Disk (B) Computer: IBM PC Compatible Computer

(〇 Operating system: P C-D O S / M S-D O S (Π Body: P a t e n t I n R e 1 e a s e # 1 · 0, V e r s i ο η # 1, 2 5 (vi) current application data

(A) Application number: US (B) Application date: August 13, 1994 (Class 0 (vih) Lawyer / agent information: ㈧ Name: He Gen, Qiang W. (Η 〇gan, J ohn W ·) — — — _- 47-____ This paper size applies the Chinese National Standard (CNS) Α4 size (2KΓχ297 mm) (Please read the precautions on the back before filling this page) Printed by the Consumer Affairs Cooperative of the Central Procurement Bureau of the Ministry of Economic Affairs System A7 B7 V. Description of the invention (45) (B) Registration number: 32,703 (0 Reference / File number: 3 2,4 5 8 (ix) Correspondence information: (A) Telephone: 201-83 BU 3195 (B) t #: 2 0 1-8 3 1-3 3 0 5 ⑵ SEQID Ν Ο: 1 Data: (i) Sequence characteristics: ㈧ Length: 2 2 base pairs (B) Type: Nucleic acid (C) Strand type : Single strand (D) topology: linear (ii) molecular type: D Ν A (genome) ⑽ hypothesis: none (iv) antisense: none (xi) sequence description SEQID Ν Ο: 1 __ CCTC AGGGCA GCTGCCTGCA GG (2) SEQID Ν Ο: 2 Data: (i) Sequence characteristics: ㈧ Length: 30 base pairs (B) Type: Nucleic acid (C) Strand type: Single-strand ⑽ Topology: Linear (ii) Molecular type State: D Ν Α (base Reason. Group) (Free) Hypothesis: None _-48- This paper size applies to Chinese National Standard (CNS) A4 size (210X297 mm) (please read the precautions on the back before filling this page) Printed by A7 B7, Consumer Cooperatives of the Standards Bureau. 5. Description of the invention (46) (iv) Antisense: None (xi) Sequence description SEQID Ν Ο: 2: GCTCTGACGC ATTTCTACAA CCACGACTCC ⑵ SEQID Ν Ο: 3 Information: (i) Sequence characteristics : ㈧ Length: 49 base pairs (B) Type: Nucleic acid (C) Strand type: Single strand ⑽ Topology: Linear ⑻ Molecular type: D Ν A (Genome) ⑽ Hypothesis: None (iv) Antisense : None (xi) SEQ ID NO: 3: TAATCCTGCA GGCAGCTGCC CTGAGGATCA GCAACTATAT ATTAAGCCG ⑵ SEQIDN 0 · 4 Information-· (i) Sequence characteristics: ㈧ Length: 46 base pairs (B) Type: Nucleic acid (0 strands) Type: Single-stranded pseudotopology: Linear (ii) Molecular type: D Ν A (Genome) (free) Hypothesis: None (iv) Antisense: None (xi) Sequence description SEQID Ν Ο: 3: _-49 -_ This paper size applies to China National Standard (CNS) A4 (210X 297mm) (Please read the back first Note: Please fill in this page again.) Printed by A7 B7 of the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs. 5. Description of the invention (47) GCTGCCTGCA GGATTATGTA AATAATTAAA ATGATAACCA TCTCGC ⑵ SEQID Ν Ο: 5 Information: (i) Sequence characteristics: (A) Length: 22 base pairs, ⑽ form: nucleic acid (0 strand type: single strand (D) topology: linear (ii) molecular form: DN A (genome) '㈣ hypothesis: none (iv) antisense: none (xi) Sequence description SEQID ΝΟ · 5: GGATTTCCTT GAAGAGAGTG AG ⑵ SEQIDN 0: 6 Materials: (i) Sequence characteristics: (A) Length: 100 base pairs ㈣ Form: nucleic acid. (0 strand type: Single strand (D) topology: linear (ii) molecular form: D Ν A (genome) (ih) hypothesis: none (iv) antisense: none (xi) sequence description SEQID Ν Ο: 6: AGCCCCCGAG TGCCTGCTCT CGAACTATTG CAACAATGAA TGCACCAAGG TGCACTACGC 60 TGACAAGGGC TACTGTTGCC TTCTGTCCTG CTATTGCXTC 100 _-JOj-_ This paper size applies to China National Standard (CNS) A4 size (210X297 mm) nnn In ϋ m 1 _ n-^ nm Ii nnm T ♦., V ' Mouth (please read the notes on the back first (This page) A7 B7 printed by the Consumer Cooperative of the Central Standards Bureau of the Ministry of Economic Affairs 5. Description of the invention (48) ⑵ SEQIDN 0: 7 Information: ① Sequence characteristics: ㈧ Length: 1 1 0 base pairs ⑽ Pattern: Nucleic acid (0 Strand type: single-stranded topology: linear · (ii) molecular type: DN A (genome) (free) hypothesis: none (iv) antisense: none (xi) sequence description SEQID Ν Ο: 3: CXGTAGGTAC CGGATCCTXA GTTAATGATG GTGGTGTCAC AGTAGCTCTT GCGAGTATCA 60 GAGATTTCCA GAACTTTCTT GTCGTCGTTG AGACCGAAGC AATAGCAGGA 110 __________ I ^ __,-/ (Please read the precautions on the back before filling out this page), ya printed by A7 B7, the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs, 5. )

AGGCACTCGG GGGCTTTTCC GGATGAGGTC GACTGCGTAG CCGTTCTTCT T ⑵ SEQID Ν Ο: 9 Data: (i) Sequence characteristics: (A) Length: 84 base pairs (B) Type: Nucleic acid (0-strand type: Single-strand ⑽ Topology: Linear ( ii) Molecular form: D Ν A (Genome) (free) Hypothesis: None (iv) Antisense: None (xi) Sequence description SEQIDN 0: 9: CCCCCCGGAT CCATGTACAA ACTGACCGTC TTCCTGATGT TCATCGCCTT CGTGATTATC 60 GCTGAGGCCA AGAAGAACGG CTAC 84 (2) SEQID Ν O: 1 0 Data:, (i) Sequence characteristics: ㈧ Length: 7 5 base pairs (B) Type: Nucleic acid (0-strand type: Single-strand ⑽ Topology: Linear (ii) Molecular type: D Ν A (Genome) ® Hypothesis: None (iv) Antisense: None _152: _ This paper size applies Chinese National Standard (CNS) A4 specification (210X 297 mm) (Please read the precautions on the back before (Fill in this page) -5 A7 B7 V. Description of the invention (50) (xi) Sequence description SEQIDN 〇: 1 0: CCCCCCGGAT CCATGTTCAA GTTCGTGATG ATCTGCGCCG TCCTCGGCCT GGCTGTGGCC 60 AAGAAGAACG GCTAC 75 (2) SEQ ID ΝΟ: 11 Information: (i) Sequence Characteristics: (A) Length: 27 bases For (B) type: Nucleic acid '(0-strand type: Single-stranded ⑽ Topology: Linear (ii) Molecular type: DNA (Genome) Inter-hypothesis: None (iv) Antisense: None (xi) Sequence description SEQIDN Ο: 1 1:

ATGTACAAAC TGACCGTCTT CCTGATG ^^^ 1- HHB vm n ^ i fl ^ in ^^^^ 1 mu nf ^ ^^^^ 1 1 ^ 1 ^ 1 nv., ·, K ,! (Please read the notes on the back before filling out this page) Printed by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Economic Affairs -53- This paper size applies to China National Standard (CNS) A4 (210 X 297 mm)

Claims (1)

Printed by the Consumers' Cooperative of the Central Bureau of Standards, Ministry of Economic Affairs • A method for preparing coated insecticides, which includes: a) Plastics that vary according to p 及 and plasticization as needed ^ Just made; Kun He B) adjust the ρΗ of the mixture in step (a) to a value above ρ η that allows the polymer to dissolve in accordance with η ′, to dissolve the polymer that changes in ρ Η c) in step (b) Insecticides, diphenylethyl compounds to be used in the UV protective gas, shanyueyue dissolving agent and sliding agent, if necessary, are added to the solution, and blended to form a homogeneous suspension. 夂 d) Make the step ( c) drying the homogeneous suspension; and if necessary e) the dry substance in the grinding step (d), wherein the polymer whose pH varies is selected from the group consisting of methyl malonic acid and formazan 84 丨 06935 patent application g Chinese Amendment to the scope of patent application (November 1987) "L / o Patent scope of water application methyl acrylate copolymer, methacrylic acid and methacryl methacrylic acid and polymer mixture, maleic anhydride and phenethyl Associative copolymer; and, pH adjustment in v (b) pH 8.5 to 10. The 2. The method according to item 1 of the scope of patent application, wherein the plasticization is # W 1 糸 selected from. Polyethylene glycol, polypropylene glycol 'g diacid diethyl ether, g dibutyl diacid ^ j Sample, S purpose, blue sesame oil, and glycerol triacetate purpose; The test system selects ammonium, alkali metal hydroxide, and alkaline earth metal hydroxide .... 'milk, read the pesticide as an insecticidal pathogen The ultraviolet protective agent is selected from the group consisting of carbon black, benzophenone, dyes and titanium dioxide; the disintegrant is selected from the group consisting of the condensation products of the sulfonation products of methyl Yue and polycyclic aromatic compounds, hydrophilic starch, carboxymethyl Cellulose and polyvinylpyrrolidine; and the sliding agent is selected from the group consisting of talc, magnesium stearate, calcium stearate, and calcium sulphate 3 The paper size is applicable to Chinese National Standard (CNS) A4 (210 × 297 mm) ..---- ^ 装 ------ Order ------ 1 ί Please read the notes on the back before filling out this page} Printed by the Employees' Cooperatives of the Central Procurement Bureau of the Ministry of Economic Affairs A method for applying an insecticide, which includes: a) a polymer which is changed according to p Η and plasticized as needed ^ Properties; kun compound; b) adjust the ρΗ of the mixture in step (a) so that the polymer dissolves according to η, .., 孓 above ρ 'to dissolve the polymer that changes according to ρ Η c) in Step (b) Add insecticide, stilbene compound to be used in the ultraviolet shielding gas, Shan Yueyue decomposer if necessary and slip agent if needed, and mix to form a homogeneous suspension. 夂 d ) Drying the homogeneous suspension in step (c); and e) grinding the dried material in step (d) if necessary, wherein the polymer whose pH varies is selected from the group consisting of methyl malonic acid and formazan 84 丨 06935 Application g Chinese Patent Application Amendment (November 1987) "L / o Patent Application Scope Water Requirement Methyl Acrylate Copolymer, Methacrylic Acid and Methacrylic Acetyl Acetate and Polymer Mixture and Malay Copolymer of acid anhydride and phenylethylene; and, the pH in v (b) is adjusted to pH 8.5 to 10. 2. The method according to item 1 of the scope of patent application, wherein the plasticization is # W 1 糸 selected from. Polyethylene glycol, polypropylene glycol 'g diacid diethyl ether, g dibutyl diacid ^ j Sample, S purpose, blue sesame oil, and glycerol triacetate purpose; The test system selects ammonium, alkali metal hydroxide, and alkaline earth metal hydroxide .... 'milk, read the pesticide as an insecticidal pathogen The ultraviolet protective agent is selected from the group consisting of carbon black, benzophenone, dyes and titanium dioxide; the disintegrant is selected from the group consisting of the condensation products of the sulfonation products of methyl Yue and polycyclic aromatic compounds, hydrophilic starch, carboxymethyl Cellulose and polyvinylpyrrolidine; and the sliding agent is selected from the group consisting of talc, magnesium stearate, calcium stearate, and calcium sulphate 3 The paper size is applicable to Chinese National Standard (CNS) A4 (210 × 297 mm) ..---- ^ 装 ------ Order ------ 1 ί Please read the precautions on the back before filling out this page} A8 B8 C8 D8 VI. Applications for patents Polymers are selected from: Copolymer of methacrylic acid and methyl methacrylate, mixture of copolymer of methacrylic acid and methyl methacrylate, and horse Copolymer of acid anhydride and phenethylene; the insecticide is an insecticidal pathogen; the plasticizer is selected from the group consisting of polyethylene glycol, polypropylene glycol 'diethyl phthalate, dibutyl phthalate, citrate , Castor oil and triacetin; the ultraviolet protective agent is selected from the group consisting of: carbon black, benzophenone, dyes and dioxide; the disintegrant is selected from the group consisting of formaldehyde and polycyclic aromatic compounds sulfonated products The salt of the condensation product, hydrophilic starch, chitomethyl cellulose, and polyethylene σ bilo bite; the slipping agent is selected from the group consisting of talc, stearic acid ball, stearin, and more acid, and (^- (^ Alcohol is isopropanol. 7. A coated insecticide that contains the insecticide core, surrounded by the Consumer Cooperatives of the Central Bureau of Standards of the Ministry of Matrix Economy, the Consumer Cooperative, Yin Changnn I—--I— .1 — i-IIIII -1-111 I--_ m 3 'Instructions (please read the precautions on the back before filling in this X) Enveloping' matrix contains 2 to 25% by weight of polymer that changes with pH, 0 to 5% by weight % Plasticizer, 5 to 45% by weight UV protection agent, 0 to 75% by weight stilbene compound, 0 to 10% by weight disintegrant And 0 to 10% by weight of the sliding agent, wherein the polymer according to p Η is selected from the group consisting of: methacrylic acid and methyl methacrylate copolymers, methacrylic acid and methacrylic acid methyl ester copolymers Blends and copolymers of maleic acid and ethylene. 8. Coated insecticides according to item 7 of the scope of patent application, wherein the ratio of insecticide to substrate is 1: 1 to 1: 1. 9. The coated insecticide according to item 7 of the scope of patent application, wherein the insecticide is an insecticidal pathogen; the polymer that varies according to ρ Η is selected from the group consisting of a copolymer of methacrylic acid and methyl methacrylate, Acrylic acid and methyl-3-This paper size is in accordance with China National Standards (CNS) A4 (210 × 297 mm) Printed by the Consumer Cooperatives of the Central Standards Bureau of the Ministry of Economic Affairs A8 B8 C8 ----- D8 VI. Application Scope of patent: Methyl acrylate copolymer mixture and maleic anhydride and styrene copolymer '· The plasticizer is selected from the group consisting of: polyethylene glycol' polypropylene glycol, diethyl phthalate, dibutyl urate, citrate , Blue sesame oil and triglyceride Carbon black, benzophenone, dyes and dioxide '; the disintegrant is selected from the group consisting of salts of condensation products of formaldehyde and polycyclic aromatic compounds acidified products, hydrophilic powder, polymethyl cellulose and polyethylene Fipyridine; The sliding agent is selected from the group consisting of talc, stearic acid ball, stearic acid # 5 and sulfuric acid. 10. A wettable powder insecticidal composition comprising 2 to 25% by weight of a wetting agent; 2 to 40% by weight of a disintegrant; 10 to 70% by weight of a swelling agent; 1 to 10% by weight % Flow enhancer; 0 to 20% by weight p Η modifier; and 5 to 75% by weight coated insecticide containing an insecticide core surrounded by a matrix that contains 2 to 25% by weight % Of polymer varying depending on ρ '0 to 5% by weight of plasticizer, 5 to 45% by weight of UV protection agent 〇 to 75% by weight of stilbene compound, 0 to 10% by weight. / ◦ disintegrating agent, and 0 to 10% by weight of the sliding agent, wherein the polymer that varies according to ρ 选自 is selected from the group consisting of methacrylic acid and methyl methacrylic acid copolymer, methacrylic acid and methacrylic acid Mixtures of methyl ester copolymers and copolymers of maleic anhydride and styrene. -4-This paper size applies to Chinese National Standard (CNS) Α4 gauge (210 X 297 mm) ------ ^ ---- ^ ------. 11 ------ --Λ (Please read the notes on the back before filling this page)