Background technology
Aborning, use in a large number chemical bactericide in order to guarantee agricultural output and quality, ecotope is produced to immense pressure, and cause some pathogen to develop immunity to drugs rapidly, further strengthened the bactericide consumption, thereby cause Practice for Pesticide Residue in Agricultural Products amount severe overweight, people and animals, environment are caused to significant damage.The close attention of the public to food security; the day by day gradually high cry of global protection environment; require the scientific research personnel to find effectively and the method for preventing and treating of safety, so development and utilization and environment have compatibility, the agricultural chemicals resource of humans are subject to day by day to the favor of various countries.
Copper rosinate is a kind of novel organic copper preparation sterilization pesticide of high-efficiency low-toxicity wide spectrum, and in recent years, the organic copper preparation became " unexpected rival " who is a dark horse in disinfectant use in agriculture.The organic copper preparation comes into one's own, be by no means accidental, it than Inorganic Copper preparation have more qualitatively, the technical and advantage used: the organic copper preparation is more safe and reliable, good with Environmental compatibility, and crop is difficult for producing poisoning, and peasant's medication is felt more relieved; The organic copper preparation is difficult for causing the propagation of two spotted spider mite and blister mite because not injuring natural enemy " crinosity bacterium ", alleviated peasant's financial burden and medication hidden danger; Organic copper preparation compatibility is fairly good, can be mixed mutually and can not cause chemical reaction with most insecticide, miticide, bactericide and plant growth regulator, alleviated the work burden that the peasant causes because repeatedly spraying medicine, saved working time, medication is saved worry more; The organic copper preparation scope of application and more extensive on the operating period also is difficult for producing poisoning under florescence and young fruit period regular service conditions; The copper content of organic copper preparation is generally lower, and residual quantity is lower, and environmental pollution reduces; In the organic copper preparation, copper ion and compound synergy, the dual mechanism sterilization, sterilization is more thoroughly more efficient.The copper rosinate that class belongs to organic copper is a kind of novel organic copper preparation sterilization pesticide of high-efficiency low-toxicity wide spectrum; there is long, easy to use new feature of lasting period; many shortcomings of former bordeaux mixture have been overcome; be the desirable bactericide that replaces bordeaux mixture, and prevention protection and treatment double action are arranged.
Biogenic pesticide also claims " biopesticide ", refer to the agricultural chemicals that utilizes the living resources exploitation, its active ingredient is that nature exists, there is low toxicity, pollution-free, selectivity strong, be easy to degraded, in ecotope the accumulation toxicity possibility little, there is the characteristic such as Environmental compatibility.The ever-increasing organic food of demand, pollution-free food being had to guarantee effect, avoid or alleviated the negative effect that the applied chemistry agricultural chemicals causes, and having the advantages such as the pathogen of making is difficult to develop immunity to drugs, is desirable agricultural chemicals type.Eugenol is by leaf, stem, the root absorption of plant, and oriented upper conduction function and safety, environmental protection noresidue, and medicine effect for treating is rapid, and the lasting period is long.After the crop spraying medicine of having fallen ill, the bacterium spore is modification at once, dissolved disappearance.
Biogenic pesticide eugenol and organic copper reagent copper rosinate are composite be have not been reported for preventing and treating downy mildew of garpe.
Downy mildew of garpe main harm blade, also can infect the tender tissues such as the tender tip, inflorescence, young fruit.Downy mildew of garpe almost all has distribution in each viny region of China, mostly occurs more area and time at rainwater, in 5~June, starts to occur.In the popular time of disease, sick leaf scorch is withered caducous, sick tip distortion, and depauperation, very large to tree vigo(u)r and yield effect, be one of Major Diseases on grape.Now, in production, for the agricultural chemicals of preventing and treating downy mildew of garpe, be mainly chemical pesticide, the strong biogenic bactericide application of low toxicity, low-residual, selectivity is also seldom.The present invention is by Toxicity Determination and field trial biogenic pesticide eugenol and the independent use of chemical agent copper rosinate and the composite rear effect to downy mildew of garpe, established the optimal proportion of two kinds of agricultural chemicals, for production and application, to reach, reduce the purpose that Pesticide environment pollutes, controls germ pesticide resistance genesis.
Embodiment
Below provide the specific embodiment of the present invention, be used for formation of the present invention is further described.
Application example
One, indoor bioassay test
(1) test method
1 experimental condition
Experiment target is designated as downy mildew of garpe (Plasmopara viticola), and sample picks up from field.
2 instrument and equipments
Instrument and equipment mainly contains electronic balance, spraying apparatus, climatic cabinate, bio-incubator, culture dish, pipettor etc.
3 experimental scheme
3.1 the examination material is prepared
Select grape susceptible variety ruby, 4~6 leaf positions from up to down, clip same area, the blade that growing way is consistent, be placed in culture dish with wet cotton balls parcel petiole, and moisturizing is standby.
3.2 test medicine
The former medicine of 35% copper rosinate, Zhengzhou China Tech green science and technology Co., Ltd product; 98% eugenol, Vad, the Community of Jin Tan County city chemical Co., Ltd of dragon product; The former medicine of 98% dimethomorph, ShenYang, Liaoning Province good harvest agricultural chemicals Co., Ltd product; 4.2% eugenol copper rosinate composite bactericide, the preparation of academy of agricultural sciences, Yantai City.
3.3 test procedure
3.3.1 sporangia suspension preparation
Get the sick leaf of new fresh grape of field natural occurrence, rinse the sporangium on clean surface with running water after, be placed in moisturizing 24h under 22 ℃ of conditions, cultivate fresh sporangium.With clean writing brush, fresh sporangium is brushed in sterile distilled water, be made into sporangia suspension.Adjust under the microscope sporangia suspension concentration to 5 * 10
5individual/mL is for test determination.
3.3.2 medicament preparation
Severally treat that the reagent agent is dissolved, be made into the 10000mg/L mother liquor standby.Each eugenol and copper rosinate compound proportion all are diluted to 5~7 concentration gradients.
3.3.3 chemicals treatment
Liquid is evenly sprayed in vacuum side of blade, after the liquid natural air drying, each is processed to blade and blade and go up dorsad, by being emitted in moisture preservation box after marks for treatment.Test is not established and is made blank containing the processing of medicament.
3.3.4 leaf dish method (Schwinn etc., 1982)
The strong leaf that with card punch, will spray medicament breaks into the leaf dish of diameter 1cm, is put in the culture dish containing wet filter paper, and 10 leaf dishes of every ware, every processing triplicate, be inoculated in leaf dish back side central authorities with ready fresh sporangia suspension drop 10 μ L.On the inoculation bonnet, the ware lid, be placed in phytotron, and at continuous illumination/dark 12h alternately, cultivation temperature is 22 ℃, 5~7d experimental investigation result.
4 investigation methods and computational methods
Investigate the incidence of each processing according to leaf dish severity Scaling method.State of an illness investigation adopts 6 grades of Recording Methods of State Standard of the People's Republic of China-field efficacy experiment criterion (two), and the percentage that accounts for the leaf disc area according to lesion area is divided corresponding sick level, calculates disease index and preventive effect.
5 statistical analyses
Do regression analysis according to each drug concentration logarithm value and corresponding preventive effect probability value, calculate the EC of each medicament
50deng numerical value.Calculate the coefficient of synergism (SR) of mixture according to the Wadley method, estimate the synergy type of mixture.Estimate the synergistic effect of pharmacy mix according to coefficient of synergism (SR), SR<0.5 is antagonism, and 0.5≤SR≤1.5 are summation action, SR > 1.5 for synergistic effect.Coefficient of synergism (SR) calculates by following formula:
In formula: X
1---the EC of mixture
50theoretical value, unit is mg/L; P
a---the percentage composition of A in mixture, unit is %; P
b---the percentage composition of B in mixture, unit is %; The EC of A in the A---mixture
50value, unit is mg/L; The EC of B in the B---mixture
50value, unit is mg/L;
In formula: the coefficient of synergism of SR---mixture; X
1---mixture EC
50theoretical value, the mg/L of unit; X
2---mixture EC
50measured value, the mg/L of unit.
(2) result of the test
Adopt leaf dish method to estimate eugenol and two kinds of single doses of copper rosinate and different proportioning mixture thereof the control efficiency to downy mildew of garpe, result shows that two kinds of single doses are indoor Plasmopara viticola is all shown to good inhibition, EC
50be respectively 0.7089mg/L and 49.4991mg/L.The EC of the medicament dimethomorph commonly used of field control downy mildew
50for 5.5628mg/L.Eugenol and copper rosinate active ingredient ratio all show synergistic effect while being 1:10~40:1, and especially the synergy of 1:20 proportioning is the most remarkable, its EC
50for 6.2681mg/L, coefficient of synergism is 1.85.Illustrate, 4.2% eugenol copper rosinate composite bactericide has very high bactericidal activity to downy mildew of garpe.
The best proportioning of table 1 eugenol and copper rosinate and contrast medicament bacteriostatic activity are measured
Two, field efficacy evaluation test
The eugenol gone out according to lab screening and copper rosinate optimum compound proportion example 1:20, configured 4.2% eugenol and copper rosinate composite bactericide, wherein containing 0.2% eugenol and 4% copper rosinate.Field is with 800 times of 12% copper rosinate missible oil (EC) and 0.3% eugenol SL800 mixed, the 50% dimethomorph WP(peace gram of bucket doubly) 1500 times in contrast medicament carry out field control effectiveness test.
(1) test method
1 experimental condition
1.1 the selection of subjects, crop and kind
Subjects is downy mildew of garpe (Plasmopara viticola), and the test grape variety is Cabernet Gernischt.
1.2 crop genetics and environmental condition
The sheet trellis type cultivation of test garden, age of tree life in 7 years, downy mildew over the years occurs general and even.Duration of test (25~August 20 July) grape breeding time is the young sprout pumping phase.The cultivation condition of all experimental plots (soil types, fertilising, growing stage, seeding row spacing etc.) and tree body growing way homogeneous, and consistent with local cultivation step.
2 experimental scheme and arrangement
2.1 medicament
2.1.1 test medicine
4.2% eugenol copper rosinate composite bactericide, the preparation of academy of agricultural sciences, Yantai City.
2.1.2 contrast medicament
12% copper rosinate EC, Shipule Pesticides Tech Development Co., Ltd., Tianjin City's product;
0.3% eugenol SL, Shandong Province hundred million good agriculture Co., Ltd product.
50% dimethomorph WP(peace gram), German BASF Trading Co., Ltd product
2.1.3 dosing arranges with processing
Every community grape strain tree is 10 strains, repeats 4 times.
Table 2 reagent agent experimental scheme
3 application methods
3.1 using method
Be watered spraying, after first dissolving with a small amount of water, fill up again institute's water requirement; During spraying, make leave dual sides even medicine, till dripping.
3.2 spraying equipment
Spraying equipment adopts SDR-120 type power spraye, injection diameter 0.8mm, operating pressure 2.0 bar.
3.3 spraying time and number of times
Dispenser first in early stage occurs in the field downy mildew of garpe, sprays altogether 3 times.The concrete dispenser date is on July 26th, 2013, August 3 and August 13.
3.4 use capacity
By amount of liquid medicine, be 150L/667m
2.
3.5 prevent and treat the medicament data of other damage by disease and insect
Duration of test ,Ge community, except the spraying test medicament, does not spray other any bactericide.
4 investigation, record and measuring method
4.1 investigation method, time and number of times
4.1.1 control time and number of times
Before medicine without downy mildew of garpe, the 8d(8 month 21 after dispenser finishes) investigate drug effect one time.
4.1.2 investigation method
20 young sprouts of every community investigation, every tip is investigated whole blades from top to bottom, puts down in writing respectively the sick numbers of sheets at different levels.Blade severity Scaling method: 0 grade: without scab; 1 grade: lesion area accounts for below 5% of whole leaf area; 3 grades: lesion area accounts for 6%~5% of whole leaf area; 5 grades: lesion area accounts for 26%~50% of whole leaf area; 7 grades: lesion area accounts for 51%~75% of whole leaf area; 9 grades: lesion area accounts for more than 76% of whole leaf area.
4.1.3 drug effect computational methods
4.2 the direct impact on crop
Test medicine, contrast medicament are to confession examination grape variety safety, without poisoning.
(2) result of the test
4.2% 800,1000,1200 times of eugenol copper rosinate composite bactericides all show good preventive effect, preventive effect is all more than 80%, be respectively 90.61%, 86.49%, 80.08%, wherein on 800 times of liquid 5% levels of composite bactericide, be significantly higher than 50% dimethomorph WP(peace gram) 1500 times, with 12% copper rosinate EC800 doubly and 0.3% eugenol SL800 doubly the mixed preventive effect of bucket there is no difference, 1000 times of liquid preventive effects of composite bactericide are 86.49%, preventive effect with contrast medicament 50% dimethomorph WP(peace gram) 1500 times there is no significant difference.Absolutely prove, 4.2% eugenol copper rosinate composite bactericide is stronger to the bactericidal activity of downy mildew of garpe, and its control efficiency and chemical pesticide are suitable, but the part substituting chemical pesticide.And 800,1000,1200 times of composite bactericides have reduced by 50.00%, 60.00% and 66.67% pesticidal preparations consumption with the bucket mixed phase than respectively, have increased the selectivity of preventing and treating the downy mildew of garpe medicament, have broad application prospects and develop value.During this, investigate, each is processed to supplying examination host plant safety, without poisoning.
Table 34.2% eugenol copper rosinate compositional bactericidal agent for preventing and treating downy mildew of garpe field control effectiveness test is table as a result
Annotate: in table, data are and manage average data everywhere
Three, mixture safety evaluatio
In complex composition, eugenol and copper rosinate content are less than the content of commercially available prod, so its toxicity to people and animals is satisfactory with the safety to crop.Composite bactericide shows to belong to the micro-virus kind agricultural chemicals through assert mechanism's toxicity test result:
Rat acute Oral toxicity LD50 ♂ > 5000mg/kg, ♀ > 5000mg/kg;
Rat acute percutaneous toxicity LD50 ♂ > 5000mg/kg, ♀ > 5000mg/kg;
It is 0(1h that lagophthalmos stimulates integration), after 48h, be 0(1:50) dilution, to the eye nonirritant;
It is 0(4h that lagophthalmos stimulates integration), to the skin nonirritant.