A kind of seed treatment containing bacillus subtilis and its production and use
Technical field
The present invention relates to a kind of seed treatment, particularly a kind of seed treatment containing bacillus subtilis and ethiprole and preparation method thereof and associated uses.The invention belongs to technical field of pesticide.
Background technology
In modern agricultural production, utilize to seed, kind of clothing inorganic agent carries out that chemicals treatment is that control soil insect and crops soil pass, one of the method for the economical and convenient of seed-borne disease.Seed treatment is a kind of for crop or other plant seed treatment, the pesticidal preparations with film forming characteristics, normally germinate required moisture and air of seed is allowed to pass through, the material slow releasing such as contained agricultural chemicals and seed manure, can kill soil insect, seedling stage insect, there is control sprout term disease and systemic disease, improve percentage of seedgermination, promote the effects such as seedling healthy growth.
Suspension seed-coating agent is the one of seed treatment, is a kind of flowable stable unit for uniform suspension be made up through wet pulverizing of pesticide original medicine, film forming agent, wetting agent, dispersant, thickener, colouring agent, filler and water.Because it is easy to use, safe, receive much attention in the seed coat agent of numerous kinds.
At present, the demand of the annual crop seeds of China is about 7,000,000 tons, wherein commercial seed amount about 3,500,000 tons, and Cotton seeds accounts for about 10%, and the market demand of visible two kinds of seed coating medicines is huge, and prospect is very wide.
Summary of the invention
The object of the invention is the seed treatment in order to provide a kind of pesticide-germicide.
Based on this purpose, the technical solution used in the present invention is:
A kind of seed treatment, its effective ingredient is ethiprole (Fipronil) and bacillus subtilis (Brevibacterium).
In the present invention, preferably, in described seed treatment the content of bacillus subtilis be 20 ~ 2,000 hundred million spores/gram, by weight percentage, the content of ethiprole is 1 ~ 20%.
In one embodiment of the invention, described seed treatment is Waterless Seed Dressing agent, containing bacillus subtilis 20 ~ 2,000 hundred million spore/restraint agent, by weight percentage, also containing ethiprole 1 ~ 20%, wetting agent 2.0 ~ 6.0%, dispersant 2.0 ~ 8.0%, film forming agent 0.5 ~ 4.0%, all the other supply 100% by filler.
Preferably, the wetting agent in described Waterless Seed Dressing agent is lauryl sodium sulfate, neopelex, pull open in powder or wetting and penetrating agent T one or more; Described dispersant is one or more in the formaldehyde condensation products (MF) of sodium lignin sulfonate, di-2-ethylhexylphosphine oxide bitter edible plant sodium sulfonate (NNO), naphthalenesulfonate formaldehyde condensation compound or methyl naphthalene sulfonic acid sodium.
Preferably, the film forming agent in described Waterless Seed Dressing agent is one or more in sodium cellulose glycolate, CMC or polyvinyl alcohol.
Preferably, the filler in described Waterless Seed Dressing agent is one or more in diatomite, bentonite, white carbon, fine particle calcium carbonate, kaolin or potter's clay.
In another embodiment of the invention, described seed treatment is suspension seed-coating agent, containing bacillus subtilis 20 ~ 2,000 hundred million spore/restraint agent, by weight percentage, also containing ethiprole 1 ~ 20%, wetting agent 2.0 ~ 6.0%, dispersant 2.0 ~ 8.0%, film forming agent 0.5 ~ 4.0%, warning colouration 0.5 ~ 4.0%, thickener 0.1 ~ 0.6%, antifreezing agent 1.0 ~ 4.0%, preservative 0.1 ~ 1.0%, defoamer 0.1 ~ 1.0%, surplus is distilled water.
Preferably, in described suspension seed-coating agent, film forming agent is sodium carboxymethylcellulose, sodium hydroxyethlcellulose, one or more in carboxy-propyl cellulose sodium, acrylic acid sodium, polyethylene glycol or polyvinylpyrrolidone.
Preferably, in described suspension seed-coating agent, dispersant is alkyl phenolic group polyethenoxy ether sulphonate, polycarboxylate, phosphate, sulphate, the rare ether block copolymers of polyoxyethylene polyoxy third, one or more in alkyl phenolic group polyoxyethylene groups phosphate, NNO, dispersing agent MF or fatty alcohol ethylene oxide adduct sulfonate.
Preferably, in described suspension seed-coating agent, wetting agent is one or more in alkyl phenolic group polyoxyethylene ether, lauryl alcohol polyethylene glycol oxide base ether sodium sulfate, Negel, alkylsulfonate, alkyl naphthalene formaldehyde condensate sulfonates, polyoxyethylene sulfonated hydrocarbon, fatty alcohol ethylene oxide adduct sulfonate, alkylphenol formaldehyde condensation product, ethylene oxide adduct sulfonate, alkylphenol polyoxyethylene formaldehyde condensation products or the rare ether block copolymers of polyoxyethylene polyoxy third.
Preferably, in described suspension seed-coating agent, warning colouration is one or more in acid scarlet, water-based rhodo, basic rhodamine.
Preferably, in described suspension seed-coating agent, thickener is one or more in xanthans, gelatin, gum Arabic, aluminium-magnesium silicate.
Preferably, in described suspension seed-coating agent, antifreezing agent is one or more in ethylene glycol, propane diols, glycerine, urea, ammonium sulfate, sodium chloride, calcium chloride.
In described suspension seed-coating agent, preservative is paraformaldehyde.
In described suspension seed-coating agent, defoamer is silicone defoaming agent.
Seed treatment of the present invention is with bacillus subtilis and the composite biological seed inorganic agent of ethiprole two kinds of effective ingredients, there is the feature efficient, safety, environment compatibility are good, energy pesticide-germicide, can be used for preventing and treating the soil insect of harm seed, vegetative period sucking pest and plurality of plant diseases, be conducive to realizing seed standardization, meet the requirement of modern agriculture with high efficiency.
Bacillus subtilis is a kind of novel microbial source biopesticide.The competition in nutrition and site, space can be formed with other harmful bacteria, can produce and there is surfactin that is antibacterial, antiviral and biosurfactant effect.The secondary metabolites of producing bacillus subtilis life produces dissolution to other harmful pathogen archespore walls, makes the cell wall perforation of harmful levels of pathogens, deformity etc.In addition, bacillus subtilis, can also Promoting plant growth as microorganism mushroom.
Ethiprole is a kind of Phenylpyrazole insecticides, not only can be used for seed treatment but also can carry out foliar spray, insecticidal spectrum is wide, to insect based on stomach poison function, have concurrently and to tag and certain systemic action, it imposes on soil can effectively chrysomelid, the wireworm of control of maize root and cutworm.
Bacillus subtilis and ethiprole compatibility do seed treatment; both are mutually collaborative; can act on from two different targets simultaneously; for seed provides omnibearing protection; its Environmental compatibility is good; toxicity is low, stores, the bio-safety such as natural enemy to people, belongs to the biological seed inorganic agent of broad-spectrum high efficacy.This seed treatment can be made into Waterless Seed Dressing agent, suspension seed-coating agent, by seed dressing or seed pelleting, for soil insects such as crop grub, wireworm, mole cricket such as control of maize, wheat, paddy rice; Also can play preventive and therapeutic effect to multiple diseases such as the fusarium wilt of crop, banded sclerotial blight, full rot, rice blast, gray mold, powdery mildew, bacterial wilts simultaneously.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage and disadvantage of the present invention will be more clear along with description.But these embodiments are only exemplary, do not form any restriction to scope of the present invention.It will be understood by those skilled in the art that and can modify to the details of technical solution of the present invention and form or replace down without departing from the spirit and scope of the present invention, but these amendments and replacement all fall within the scope of protection of the present invention.
The preparation of embodiment 1, Waterless Seed Dressing agent
Embodiment 1-1
Bacillus subtilis starter medicine (1,000,000,000,000 spores/gram) 0.2kg, ethiprole (folding hundred) 1kg, wetting agent pull open powder 2.0kg, dispersant NNO4.0kg, film forming agent CMC 0.5kg, another kaolin supplies 100kg, through air-flow crushing after each component mixes, can be made into the Waterless Seed Dressing agent 100kg of 1% ethiprole 2,000,000,000 living spores/gram bacillus subtilis.
Embodiment 1-2
Bacillus subtilis starter medicine (1,000,000,000,000 spores/gram) 10kg, ethiprole (folding hundred) 10kg, wetting agent lauryl sodium sulfate 4.0kg, dispersant sodium lignin sulfonate 6.0kg, film forming agent sodium cellulose glycolate 2.0kg, another kaolin supplies 100kg, through air-flow crushing after each component mixes, can be made into the Waterless Seed Dressing agent 100kg of 10% ethiprole 1,000 hundred million living spores/gram bacillus subtilis.
Embodiment 1-3
Bacillus subtilis starter medicine (1,000,000,000,000 spores/gram) 20kg, ethiprole (folding hundred) 20kg, wetting agent neopelex 4.0kg, dispersant sodium lignin sulfonate 6.0kg, film forming agent sodium cellulose glycolate 2.0kg, another fine particle calcium carbonate supplies 100kg, through air-flow crushing after each component mixes, can be made into the Waterless Seed Dressing agent 100kg of 20% ethiprole 2,000 hundred million living spores/gram bacillus subtilis.
The preparation of embodiment 2, suspension seed-coating agent
Embodiment 2-1
Bacillus subtilis starter medicine (1,000,000,000,000 spores/gram) 0.2kg, ethiprole (folding hundred) 1kg, wetting agent lauryl alcohol polyethylene glycol oxide base ether sodium sulfate 3.0kg, dispersant alkyl phenolic group polyoxyethylene groups phosphate 6.0kg, film forming agent sodium cellulose glycolate 2.0kg, warning colouration is acid scarlet 1.0kg, thickener xanthans 0.2kg, antifreeze glycol 4.0kg, preservative paraformaldehyde 0.3kg, silicone defoaming agent 0.2kg, another water supplies 100kg, to mix and through ball milling, sand milling is pulverized, dispersion, can be made into the aqueous suspension agent 100kg of 1% ethiprole 2,000,000,000 living spores/g bacillus subtilis.
Embodiment 2-2
Bacillus subtilis starter medicine (1,000,000,000,000 spores/gram) 10kg, ethiprole (folding hundred) 10kg, wetting agent lauryl sodium sulfate 4.0kg, dispersant NNO6.0kg, film forming agent CMC 3.0kg, warning colouration is basic rhodamine 2.0kg, thickener xanthans 0.2kg, antifreezing agent propane diols 4.0kg, preservative paraformaldehyde 0.3kg, silicone defoaming agent 0.2kg, another water supplies 100kg, mixes and pulverizes, disperses, can be made into the aqueous suspension agent 100kg of 10% ethiprole 1,000 hundred million living spores/g bacillus subtilis through ball milling, sand milling.
Embodiment 3 effect test
One, indoor sterilization insecticidal activity assay
1.1 sterilizations (Rhizoctonia solani Kuhn) determination of activity
(1) test preparation medicament
Taking bacillus subtilis starter medicine (1,000,000,000,000 spores/g) is distributed in the 0.1% Tween 80 aqueous solution, be mixed with one group of bacillus subtilis single dose: establish 6 concentration gradients, in female medicine, concentration be respectively 4 μ g/mL(0.04 hundred million spores/gram), 8 μ g/mL(0.08 hundred million spores/gram), 16 μ g/mL(0.16 hundred million spores/gram), 32 μ g/mL(0.32 hundred million spores/gram), 64 μ g/mL(0.64 hundred million spores/gram), 128 μ g/mL(1.28 hundred million spores/gram).
Take bacillus subtilis starter medicine (1,000,000,000,000 spores/g) and ethiprole, be distributed in the Tween 80 aqueous solution of 0.1%, prepare 5 groups of ethiproles/bacillus subtilis intermixture: establish 6 concentration gradients, in group, the concentration of each dose of bacillus subtilis is that 4,8,16,32,64,128 μ g/mL(are in female medicine), in each group medicament, the addition of ethiprole and the weight ratio of bacillus subtilis starter medicine are respectively 5.1:0.4,5.05:0.45,5.0:0.5,4.95:0.55,4.9:0.6.
(2) Plating determination experiment
Each experimental pharmacy is proceeded as follows respectively: get 1mL test medicine, joins about 45 DEG C, shake up in 49mL quantitative culture medium after pour in culture dish, cool for subsequent use.By cultured pathogen, aseptically steal bacterium cake with the sterilizing card punch of diameter 5mm from colony edge, with inoculator, pure culture biscuits involvng inoculation is central in flat board, mycelia faces up, and covers culture dish, at 25 DEG C of dark culturing 4d.Each medicament carries out 4 groups of parallel tests simultaneously, and adds medium as blank with the 0.1% Tween 80 aqueous solution.
(3) investigation and statistical analysis
At control group colony growth to carrying out measurement investigation during culture dish edge.
Adopt right-angled intersection method to measure colony diameter, calculate bacteriostasis rate, pharmacy test concentration and bacteriostasis rate are carried out respectively logarithm and the conversion of probability value, with log concentration value for independent variable, bacteriostasis rate probability value is dependent variable, carries out regression analysis, obtains virulence regression equation, EC
50and EC
90correlation coefficient and coefficient of synergism, the data obtained is as shown in table 1.
According to the virulence size of these data evaluation bacillus subtilis Rhizoctonia solani and ethiprole on the impact of bactericide bactericidal effect.
1.2 desinsections (grub) determination of activity
(1) test preparation medicament
Ethiprole is distributed in the Tween 80 aqueous solution of 0.1%, prepares one group of ethiprole single dose: establish 6 concentration gradients, be respectively 5,10,20,40,80,160mg/L.
Ethiprole, bacillus subtilis starter medicine (1,000,000,000,000 spores/g) are distributed in the Tween 80 aqueous solution of 0.1%, prepare 5 groups of ethiproles and bacillus subtilis intermixture: establish 6 concentration gradients, wherein the concentration of ethiprole be respectively 5,10,20,40,80,160mg/L; The weight ratio of each group of ethiprole and bacillus subtilis starter medicine is respectively: 5.1:0.4,5.05:0.45,5.0:0.5,4.95:0.55,4.9:0.6.
(2) grub test method
Pesticide-clay mixture method is adopted to measure above each test medicine the virulence of grub respectively: dry ground earth to be crossed 20 order testing sieves, then get 800g and load (high 10cm in circular cultivation box, diameter 13cm), getting 100ml test medicine pours into wherein, mix evenly after Indoor Natural infiltration 2h, then the wheat seed showed money or valuables one carries unintentionally by imbibition is mixed into wherein; The grub selecting 15 sizes more consistent puts into box, allows its bury voluntarily (that can not bury voluntarily will be changed), then puts into the recovery room of 26 DEG C, check result after 48h.Each test medicine carries out 4 groups of parallel tests simultaneously, and is mixed in soil as blank with the Tween 80 aqueous solution of 0.1%.
(3) investigation and statistical analysis
Obviously shrink can not be considered as death by scrambler with polypide during check result.
Calculate lethality, use Abbott formula to correct lethality, then with log concentration value for independent variable, the probability value of corrected mortality is dependent variable, ask the virulence regression equation of each single dose and complex preparation, LC50, LC90, correlation coefficient and coefficient of synergism, the data obtained is in table 2.
By the virulence size of these data evaluation ethiproles to grub, analyze bacillus subtilis to the impact of ethiprole virulence size, determine the optimum proportioning of bacillus subtilis and ethiprole.
The experimental data of table 1 and table 2 shows, ethiprole has certain facilitation to bacillus subtilis activity, and bacillus subtilis also demonstrates certain synergistic effect to the insecticidal activity of ethiprole, the two reasonable compatibility, can reach the effect of disinsection prophylaxis simultaneously.
Table 1 ethiprole and bacillus subtilis compound sterilizing synergy situation
Table 2 ethiprole and bacillus subtilis composite control of grubs synergy situation
Two, field experiment
2010, experimental field, Linqing, Shandong, seed treatment of the present invention carried out to wheat seed dressing and dressing test, the results are shown in Table 3; 2011, experimental field, Potou, carry out wheat seed dressing and dressing test to seed treatment of the present invention, result of the test was in table 4.
The experimental data of table 3 and table 4 shows, seed treatment of the present invention has following characteristics:
1, by crop seed dressing or Dressing, to the multiple diseases such as banded sclerotial blight, fusarium wilt, full rot, root rot, smut of corn, wheat, good preventive effect is had to soil insect.
2, seed can be made effectively to avoid the invasion of damage by disease and insect after seed seed dressing or dressing, field lasting period length has synergistic effect.
3, said preparation effective ingredient ethiprole, bacillus subtilis, the two compatibility, safety is high, has production-increasing function, good to environment compatibility.
4, ethiprole of the present invention and bacillus subtilis complex biological seed treatment are by dress seed or dressing provides comprehensive protection to seed, meet the developing direction of seed standardized production, have larger society and economic worth.Seed treatment of the present invention not only contributes to producing free of contamination Pesticidal products, can also keep the balance of agricultural ecosystem and bio-diversity simultaneously, more be conducive to agricultural sustainable development.
Table 32010 year experimental field, Linqing, Shandong control of maize damage by disease and insect result of the test
Table 42011 year Potou experimental field of wheat control Wheat Diseases And Insect Pests result of the test