Summary of the invention
In order to reduce the use of agricultural chemicals and to improve the control efficiency to disease, we are by composite to biogenic pesticide Ningnanmycin and chemical bactericide imazalil in the present invention, by Toxicity Determination and field trial, establish the optimal proportion of two kinds of medicaments, for production and application, ring rot of apple and brown spot are had to good control efficiency, also reduce pesticide dosage simultaneously, reduce environmental pollution.
The present invention is inquiring into the application in agriculture field of imazalil and complex composition thereof, especially studied for ring rot of apple and Control of Lecanosticta Acicola, aimed to provide a kind of Ningnanmycin and imazalil single dose and complex composition thereof of preventing and treating ring rot of apple and brown spot.
The present invention is achieved by the following technical solutions:
The bactericidal composition of control apple disease, special character is, the weight proportion of its active ingredient imazalil and Ningnanmycin is 8:1~1:8;
The bactericidal composition of control apple disease, further improves and is, the synergy weight proportion of its active ingredient imazalil and Ningnanmycin is 1:2~8;
The bactericidal composition of control apple disease, further improves and is, the optimal proportion of its active ingredient imazalil and Ningnanmycin is 1:2.
The bactericidal composition of control apple disease, its active ingredient imazalil and the weight percentage of Ningnanmycin in whole complex composition are respectively 2% and 4%, and surplus is conventional auxiliary material additive.
Imazalil has good control efficiency to ring rot of apple, and Ningnanmycin has good control efficiency to apple marssonina leaf spot, and a kind of Ningnanmycin of the present invention and imazalil combination sterilization composition are applied to prevents and treats ring rot of apple and apple marssonina leaf spot.
The bactericidal composition that the present invention prevents and treats apple disease dilution 600-800 doubly lower application prevent and treat ring rot of apple and apple marssonina leaf spot has good preventive effect.
Imazalil of the present invention and Ningnanmycin complex composition have the following advantages:
1, there is synergistic effect.In the indoor experiment of the preventive effect to Botryosphaeria berengeriana f. sp, imazalil and Ningnanmycin all show synergistic effect with the proportioning of 1:2,1:4,1:8, and wherein, with the proportioning synergy maximum of 1:2, coefficient of synergism is 1.74.
2, preventive effect significantly and reduce pesticide dosage.We prepare 6% imazalil Ningnanmycin compound and have outstanding performance in field control effectiveness test.Preventing and treating in ring rot of apple test, dilute 600,800 times, two and process preventive effect all more than 85%, wherein 600 times of processing preventive effects are significantly higher than 1000 times of processing of 10% imazalil aqueous emulsion and 1000 times of processing of 8% Ningnanmycin aqua, and 800 times of processing preventive effects are suitable with imazalil single dose preventive effect; After 600 times and 800 times of compound dilutions to the preventive effect of apple marssonina leaf spot all more than 85%, process preventive effects higher than 1000 times of preventive effects of 8% Ningnanmycin aqua dilution for 600 times, 800 times of processing preventive effects are suitable with it.This explanation compound drug 6% imazalil Ningnanmycin in the situation that medicament usage amount reduces still to preventing and treating ring rot of apple and brown spot has remarkable effect.
3, high security.Imazalil is low to person poultry toxicity, and Ningnanmycin is microbial pesticide, without poisoning, environmentally safe.The complex composition that we make is safer to crop, environment and fresh fruit eater.
4, reduced the germ risk that develops immunity to drugs.As described in advantage 2, the content of complex composition imazalil and Ningnanmycin is all few than the medicament single dose of market sale, in the time of field control disease, dosage also reduces, this has significantly alleviated Botryosphaeria berengeriana f. sp and the selection pressure of brown spot to medicament, in addition, imazalil is different to the mode of action of germ with Ningnanmycin, is more difficult for germ is developed immunity to drugs after composite.
Embodiment
The present invention is elaborated with following instantiation, but limitation of the present invention absolutely not.In all proportionings, percentage is all weight percentage.
Embodiment 1
Toxicity Determination and compound proportion screening
1.1 experimental pharmacy
The former medicine of 98% imazalil, Maxime chemical company of Israel product;
The former medicine of 60% Ningnanmycin, Deqiang Biology Co., Ltd.'s product.
1.2 strains tested
Experiment target is designated as ring rot of apple (Botryosphaeria dothidea) germ bacterial strain sample, and from field, upper separation of sick fruit obtains, and the first generation bacterial strain that single cell culture purifying obtains, saves backup in 4 ℃ of refrigerators.
1.3 instrument and equipment
Instrument and equipment mainly contains electronic balance, superclean bench, bio-incubator, culture dish, pipettor etc.
1.4 experimental technique
The biomass growth rate method (Mycelium growth rate test) that adopts FAO to recommend.
1.4.1 composite reagent
Table 1 imazalil and Ningnanmycin compound proportion
Compound drug diluted concentration gradient under table 2 different proportion
1.4.2 the preparation of pastille flat board
The drug concentration gradient of setting according to 1.4.1, adds medicament in the PDA medium quantitatively dissolving, and makes pastille flat board, and so that pastille flat board is not as blank, every concentration repeats 3 times.
1.4.3 connecing bacterium cultivates
Beat and get bacterium dish along cultured colony edge with diameter 0.6 ㎝ card punch, with sterilization transfer needle, the reversion of bacterium dish is transplanted to dull and stereotyped above, be placed under 25 ℃ of conditions constant temperature culture 7 days.
1.4.4 result investigation
Measure bacterium colony expansion diameter with ruler intersection mensuration, contrast bacterium dish expansion diameter with not dispenser and compare, calculate the inhibiting rate of medicament to mycelial growth.
1.5 statistical analysis
Do regression analysis according to each drug concentration logarithm value and corresponding mycelial growth inhibition rate probability value, calculate each medicament virulence regression equation, correlation coefficient (r) and EC
50.
While carrying out medicament joint toxicity measuring, calculate the coefficient of synergism (SR) of complex composition according to Wadley method, evaluate the synergy type of complex composition.Evaluate the synergistic effect of composite reagent according to coefficient of synergism (SR), SR<0.5 is antagonism, and 0.5≤SR≤1.5 are summation action, and SR>1.5 is synergistic effect.Coefficient of synergism (SR) calculates by formula (1), (2):
In formula:
X
1----complex composition EC
50theoretical value, unit is mg/L;
P
a---the percentage composition of A in-complex composition, unit is percentage (%);
P
b---the percentage composition of B in-complex composition, unit is percentage (%);
The EC of A----A
50value, unit is mg/L;
The EC of B----B
50value, unit is mg/L.
In formula:
The coefficient of synergism of SR----complex composition;
X
1----complex composition EC
50theoretical value, unit is mg/L;
X
2----complex composition EC
50measured value, unit is mg/L.
1.6 interpretation of result
Test by indoor virulence, can show that imazalil and Ningnanmycin weight ratio 8:1,4:1,2:1,1:1 have summation action, 1:2,1:4,1:8 have synergistic effect, and 1:2 is optimum compound proportion example.
Table 3 imazalil and the screening of Ningnanmycin optimum compound proportion example
Embodiment 2
Field plot trial
Filter out imazalil according to indoor compound proportion screening experiment: Ningnanmycin=1:2 is best proportioning, we have prepared 6% imazalil Ningnanmycin compound.
2.1 compound drugs are prevented and treated ring rot of apple field trial
According to the chief editor's of the Institute for the Control of Agrochemicals of the Ministry of Agriculture,PRC the field control effectiveness test of " " pesticide field efficacy medicine test criterion (two) " the 118th part: bactericidal agent for preventing and treating ring rot of apple " design 6% imazalil Ningnanmycin compound to ring rot of apple.
2.1.1 reagent agent
10% 1000 times of imazalil aqueous emulsions, Shaanxi Mei Bang agricultural chemicals Co., Ltd product;
8% 1000 times, Ningnanmycin aqua, Huifeng bio tech ltd, Tonglu, Zhejiang Province product;
6% 600 times of imazalil Ningnanmycins, 800 times, 1000 times, the preparation of Yantai academy of agricultural sciences.
2.1.2 contrast medicament
70% 1000 times of thiophanate-methyl wetting powders, Tso Tat Co., Ltd., Japan's product.
2.1.3 Quarter Design
7 processing are established in test altogether, 4 repetitions of every processing, She28Ge community altogether, 4 of every communities strain tree.Each community randomized arrangement.Spraying equipment adopts workers and peasants 26 type high pressure power driven sprayers, with in tree crown all branch blades even medicine, till dripping.
2.1.4 control time and method
Picking time, centre 2 strains were chosen in every community, investigated whole fruits, and the total fruit of record number, sick fruit number, calculate preventive effect with diseased fruit rate; Storage period, after gathering, each fruit of processing is rejected after disease, worm, wound fruit, and the same day, dress basket was put under natural room temperature, stores latter 10 days, 20 days, 30 days once sick fruit numbers of investigation respectively, calculates preventive effect with accumulation diseased fruit rate.
2.1.5 drug effect computational methods
2.1.6 result of the test
The 6% imazalil Ningnanmycin of making on the basis of lab screening is had outstanding performance in the test of field control ring rot of apple, 600 times, 800 times preventive effects are all more than 85%, wherein 600 times of processing are significantly higher than 1000 times, 1000 times of 10% imazalil aqueous emulsions and 8% Ningnanmycin aqua, 800 times of processing and quite (detailed data is in table 4) of imazalil single dose preventive effect.
Table 43% imazalil Ningnanmycin is prevented and treated ring rot of apple field efficacy
Process |
Diseased fruit rate (%) |
Preventive effect (%) |
The significance of difference (5%) |
10% 1000 times of imazalil aqueous emulsions |
3.55 |
89.25 |
b |
8% 1000 times, Ningnanmycin aqua |
8.35 |
65.23 |
d |
6% 600 times of imazalil Ningnanmycins |
2.58 |
92.33 |
a |
6% 800 times of imazalil Ningnanmycins |
3.43 |
89.81 |
b |
6% 1000 times of imazalil Ningnanmycins |
5.15 |
84.71 |
c |
70% thiophanate-methyl WP1000 doubly |
5.60 |
83.37 |
c |
Blank |
33.69 |
? |
? |
2.2 compound drugs are prevented and treated apple marssonina leaf spot field trial
With reference to the chief editor's of the Institute for the Control of Agrochemicals of the Ministry of Agriculture,PRC the field control effectiveness test of " " pesticide field efficacy medicine test criterion (two) " the 124th part: bactericidal agent for preventing and treating alternaria leaf spot of apple " design 6% imazalil Ningnanmycin compound to apple marssonina leaf spot.
2.2.1 reagent agent
10% 1000 times of imazalil aqueous emulsions, Shaanxi Mei Bang agricultural chemicals Co., Ltd product;
8% 1000 times, Ningnanmycin aqua, Deqiang Biology Co., Ltd.'s product;
6% 600 times of imazalil Ningnanmycins, 800 times, 1000 times, the preparation of Yantai academy of agricultural sciences.
2.2.2 contrast medicament
10% 1500 times of difenoconazole water dispersible granules, are first just reaching Suzhou crop science Co., Ltd product.2.2.3 Quarter Design
7 processing are established in test altogether, 4 repetitions of every processing, She28Ge community altogether, 4 of every communities strain tree.Each community randomized arrangement.Spraying equipment adopts workers and peasants 26 type high pressure power driven sprayers, with in tree crown all branch blades even medicine, till dripping.
2.2.4 control time and method
Every community choose in the middle of 2 strains, every strain divides East, West, South, North, middlely investigates respectively 2 whole blades of branch, accounts for whole leaf area percentage carry out classification with the lesion area on every a slice leaf.Blade stage division:
0 grade: without scab;
1 grade: lesion area accounts for below 10% of whole leaf area;
3 grades: lesion area accounts for 11%~20% of whole leaf area;
5 grades: lesion area accounts for 21%~50% of whole leaf area;
7 grades: lesion area accounts for 41%~65% of whole leaf area;
9 grades: lesion area accounts for the more than 65% of whole leaf area.
2.2.5 drug effect computational methods
2.2.6 result of the test
The 6% imazalil Ningnanmycin compound of making on the basis of lab screening is had outstanding performance in the test of field control apple marssonina leaf spot, 600 times, 800 times preventive effects are all more than 85%, wherein process preventive effect and be significantly higher than 1000 times, 1000 times of 10% imazalil aqueous emulsions and 8% Ningnanmycin aqua for 600 times, 800 times of processing are suitable with Ningnanmycin preventive effect, higher than the preventive effect (detailed data is in table 5) of imazalil.
Table 56% imazalil Ningnanmycin is prevented and treated apple marssonina leaf spot field efficacy
Process |
Disease refers to |
Preventive effect |
The significance of difference (5%) |
10% 1000 times of imazalil aqueous emulsions |
13.64 |
67.47 |
e |
8% 1000 times, Ningnanmycin aqua |
6.03 |
90.04 |
c |
6% 600 times of imazalil Ningnanmycins |
4.06 |
93.30 |
b |
6% 800 times of imazalil Ningnanmycins |
7.05 |
88.35 |
c |
6% 1000 times of imazalil Ningnanmycins |
9.50 |
84.30 |
d |
10% 1500 times of difenoconazole water dispersible granules |
1.01 |
98.33 |
a |
Blank |
60.53 |
? |
? |
Embodiment 3
Safety testing
In complex composition, imazalil and Ningnanmycin content are less than the content of commercially available prod, and therefore its toxicity to people and animals and the safety of crop are satisfactory.Composite bactericide shows to belong to 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 ♂ >20000mg/kg, ♀ >20000mg/kg.