CN104542658B - A kind of composite bactericide and preparation method thereof - Google Patents
A kind of composite bactericide and preparation method thereof Download PDFInfo
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- CN104542658B CN104542658B CN201510046859.0A CN201510046859A CN104542658B CN 104542658 B CN104542658 B CN 104542658B CN 201510046859 A CN201510046859 A CN 201510046859A CN 104542658 B CN104542658 B CN 104542658B
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Abstract
The invention discloses a kind of composite bactericide and preparation method thereof, composite bactericide is made up of active component, filler and auxiliary agent, wherein, active component is made up of triazole bactericidal agent Tebuconazole, benzimidazole germicide carbendazim.Can the limb fungal disease such as effectively preventing and treating apple-tree skin rotten pathogenic bacteria, Botryosphaeria berengeriana f. sp.Present invention synergistic effect on apple tree canker fungal disease is prevented and treated is notable, and X%DW paste roots out effect up to 100% in 120 days, obviously higher than unitary agent；Complex preparation quick-acting and lasting effect are good, and mechanism of action and site are more, can effectively delay the drug-fast occurrence and development of pathogen.In addition, the preparation technology of the present invention is simple, cost is low, remarkable in economical benefits, and diseases prevention spectrum is wide, and biological safety is high.
Description
Technical field
The invention belongs to planting fruit trees technical field, more particularly to a kind of composite bactericide and preparation method thereof.
Background technology
It is a kind of distribution on apple as caused by Valsa Valsa ceratosperma that apple tree canker, which is,
Extensively, the disease of weight is endangered.Since finding apple tree canker first from China, there have been big four times prevalence (Zeng Shimai the country
Macroscopical Plant Pathology Beijing：Chinese agriculture publishing house, 2005).2007, national Apple Industry technical system was set up, and is complete
The investigation and prevention and control research of apple pest create condition in the range of state, and the investigation result of 2008 is shown, national apple master
The average onset strain rate that production saves canker of apple fruit is 52.7%, it can be seen that the generation of rot is very universal and serious.
Chemical prevention is to control one of effective measures of apple tree canker, and common fungicide has thiophanate-methyl, first sulphur
Methyl α-naphthyl acetate, humic acid ketone, ambam etc..In process of production, frequently apple tree corruption is easily produced using single chemical agent
Rotten disease causes prevention effect to drastically reduce the resistance to the action of a drug of bactericide.Therefore, new, efficient, safe chemical bactericide is found
It is urgent problem to be solved in current production.
Apple tree canker cause of disease Perfect stage is apple Valsa [Valsa mali Miyabe et Yamada],
Ascomycotina, Valsa；Invisible element be shell capsule spore [Cytospora mandshurica Miura], Deuteromycotina,
Cytospora.Tebuconazole is triazole bactericidal agent, and triazole bactericidal agent main function mechanism is destruction and prevents germ cell membrane
The biosynthesis of important composition composition ergosterol, causes cell membrane to be formed, and makes germ dead, higher fungus is shown
Obvious inhibitory action.Carbendazim also known as carbendazol, carbendazol.Carbendazim is a kind of broad-spectrum germicide, to a variety of
Crop disease as caused by fungi (such as Fungi Imperfecti, many sac fungus) has prevention effect.
In addition, liniment is to cure the most suitable formulation of rot wound, it is also the development for preventing and treating rot medicament at present
Direction.Registered medicament is generally that liniment (paste, paste, liniment) has the formulation of certain viscosity, can so stick tree
Body tissue, can prevent the intrusion of external germ, and the rotten pathogenic bacteria of residual in tissue can be killed again.Due to the anniversary of germ spore
Propagate and wound is numerous, the characteristics of new medicament must also have long-term effect and be easy to use.It can thus be appreciated that formulation is smeared in research and development
Medicament is to prevent and treat the important research direction of rot.
At present, there is not yet on carbendazim and Tebuconazole are combined into composite bactericide and the report of paste making agent is done.
The content of the invention
It is an object of the invention to provide a kind of composite bactericide and preparation method thereof, it is intended to solves existing preventing and treating and rots
The effect that the medicament of disease is present is poor, complex operation, the problem of term of validity is short.
The present invention is achieved in that a kind of composite bactericide, and the composite bactericide includes according to parts by weight：Carbendazim
2.7g, Tebuconazole 0.3g, Op10：10.20~10.70g, gibberellin GA：0.002g, 4.3 ± 0.5g of alcohol, emulsion paint polishing is extremely
100g。
Further, carbendazim, Tebuconazole are respectively active compound.
Further, composite bactericide is liniment.
Another object of the present invention is to provide a kind of composite bactericide preparation method to comprise the following steps：
2.7g carbendazim, 0.3g Tebuconazoles is taken to be put into agitator tank, plus 10.20~10.70g Emulsifier Os p10 is sufficiently stirred for
After dissolving, stand well mixed, then weigh gibberellin 0.002g with small beaker, stirring is put into after ± 0.5g alcohol dissolving of plus 4.3
Stirred in tank, emulsion paint polishing to 100g.
Further, the composite bactericide be configured to missible oil, wettable powder, granula, soluble powder, water dispersible granules,
Aqueous suspension agent, aqueous emulsion, pray for emulsion, emulsion, dry suspension, liniment.
Further, the composite bactericide application method, is smeared using limb, and sick scar scraped clean is smeared at trunk.
Composite bactericide that the present invention is provided and preparation method thereof, composite bactericide is by active component, filler and auxiliary agent group
Into, wherein, active component is made up of triazole bactericidal agent Tebuconazole, benzimidazole germicide carbendazim.Apple can effectively be prevented and treated
The apple limb fungal disease such as skin of fruit tree rotten pathogenic bacteria, Botryosphaeria berengeriana f. sp.The present invention is preventing and treating apple tree canker fungi
Significantly, X%DW paste roots out effect up to 100% to the upper synergistic effect of venereal disease evil in 120 days, obviously higher than unitary agent；Compounding
Preparation quick-acting and lasting effect are good, and mechanism of action and site are more, can effectively delay the drug-fast occurrence and development of pathogen.This
Outside, preparation technology of the invention is simple, and cost is low, remarkable in economical benefits, and diseases prevention spectrum is wide, and biological safety is high.
Brief description of the drawings
Fig. 1 is the preparation method flow chart of composite bactericide provided in an embodiment of the present invention；
Fig. 2 is that influence of the different agents to the antibacterial bandwidth of Valsa mali is shown under normal temperature provided in an embodiment of the present invention
It is intended to；
Fig. 3 is that the antibacterial band of apple bark rot is formed after 3%DW provided in an embodiment of the present invention is handled through high and low temperature
Influence schematic diagram；
Fig. 4 is that the antibacterial band of apple bark rot is formed after 5%DW provided in an embodiment of the present invention is handled through high and low temperature
Influence schematic diagram；
Fig. 5 is provided in an embodiment of the present invention to root out apple tree canker scab wound callus shape after different disposal
Into influence schematic diagram；
Fig. 6 is that different compound drugs provided in an embodiment of the present invention are determined to apple bark rotten pathogenic bacteria mycelial growth EC50
Schematic diagram；
Fig. 7 is isolated shoot experiment schematic diagram provided in an embodiment of the present invention；
Fig. 8 is the influence signal that different agents provided in an embodiment of the present invention are formed to the antibacterial band of apple rot pathogen
Figure.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
Below in conjunction with the accompanying drawings and specific embodiment to the present invention application principle be further described.
The composite bactericide of the embodiment of the present invention：It is made up of active component, filler and auxiliary agent, wherein, active component is by three
Azoles fungicide Tebuconazole, benzimidazole germicide carbendazim composition；
The composite bactericide includes according to parts by weight：Carbendazim 2.7g, Tebuconazole 0.3g, Op10：10.20~
10.70g, gibberellin GA：0.002g, 4.3 ± 0.5g of alcohol, emulsion paint polishing to 100g.
Further, carbendazim, Tebuconazole are respectively active compound.
Further, composite bactericide is liniment.
Active ingredient combinations containing enhancing effective amount；Filler is gibberellin, emulsion paint；Adjuvant used is OP10, alcohol.
Composite bactericide of the present invention, can be configured to various types of preparations, for example missible oil, wettable powder,
Granula, soluble powder, water dispersible granules, aqueous suspension agent, aqueous emulsion, pray for emulsion, emulsion, dry suspension, liniment (paste) etc..
The composite bactericide application method of the present invention, general to be smeared using limb, sick scar scraped clean mainly at trunk
Smear this bactericide.
As shown in figure 1, the preparation method of the composite bactericide of the embodiment of the present invention comprises the following steps：
S101：2.7g carbendazim, 0.3g Tebuconazoles is taken to be put into agitator tank, plus 10.20~10.70g Emulsifier Os p10 fills
Divide after stirring and dissolving, stand well mixed；
S102：Gibberellin 0.002g is weighed with small beaker again, is put into agitator tank and stirs after ± 0.5g alcohol of plus 4.3 dissolving
Uniformly, emulsion paint polishing 100g.
The application effect of the present invention is described further by following experiment：
First, materials and methods
1 test material
1.1 bacterial strains, are separated by laboratory for examination pathogen apple rot pathogen Valsa ceratosperma and preserved；
1.1.1 chemical agent, the Original Tebuconazole of triazole bactericidal agent 97%；The carbendazim of benzimidazole germicide 98% is former
Medicine；
2 methods：
Test the influence of 1 pair of apple rot pathogen mycelial growth
Determined using mycelial growth rate method, by 97% Original Tebuconazole and 98% carbendazim active compound, and Tebuconazole with
The different ratio of carbendazim is stand-by with the mother liquor that 1000ug/mL is respectively prepared after acetone solution, prepares reagent agent during experiment
Into the decoction of various concentrations, (added and filled in the PDA culture medium 99mL slightly cooled down after sterilizing with sterilizing pipette, extract 1mL decoctions
Divide and shake up, pour into culture dish, every kind of each concentration of medicament sets 4 repetitions, is compared with acetone, sterilized water makees blank control,
The bacteria cake for taking a diameter of 0.7cm is beaten in colony edge, pastille flat board center is inoculated into, 1 ferfas cake is put per ware, 25 DEG C of constant temperature trainings are put
Support in case and colony diameter (Fig. 6) is measured using crossing method after culture 4d, calculate inhibiting rate, inhibiting rate is converted into probability
Value, drug concentration is converted between log concentration, then the logarithm value for being by the niqueMin and drug concentration of relative inhibition
Linear regression analysis, obtain suppress apple rot pathogen mycelial growth 50% valid density (EC50 values), suppression
Rate %=(control mycelial growth diameter-processing mycelial growth diameter)/control mycelial growth diameter × 100%
The inhibitory action of the Tebuconazole of table 1 and carbendazim and its compounding to Valsa mali mycelial growth
Note：During SR ＜ 0.5, antagonism is represented；During 0.5≤SR≤1.5, summation action is represented；During SR ＞ 1.5, represent
Synergistic effect
As shown in Table 1, carbendazim, Tebuconazole and its different blended are determined under indoor conditions to rot to apple bark with combination
Germ mycelial growth shows different inhibitory action effects, it is determined that optimum proportioning, when carbendazim with Original Tebuconazole by 9: 1
Proportioning performance synergistic effect, coefficient of synergism is 1.533, and this proportioning is the optimum proportioning of the two.
Test the influence of 2 pairs of apple rot pathogen spore germinations：
To the inhibitory action of rotten pathogenic bacteria spore germination：The apple tree with rot pycnidia is gathered from field
Skin, at room temperature moisturizing culture promotes cirrus to produce, and picking yellow cirrus is incorporated in aqua sterilisa, and it is 1 × 10 that concentration, which is made,6~
1×107Individual conidium/mL spore suspension is standby；
Reagent agent is configured to the decoction of various concentrations during experiment, is added and sterilized with sterilizing pipette, extract 1mL decoctions
Fully shake up, poured into culture dish in the PDA culture medium 99mL slightly cooled down afterwards, every kind of each concentration of medicament sets 4 repetitions, with third
Ketone is compared, and sterilized water is made to cultivate under blank control, 25 DEG C of dark conditions, when control germination rate reaches 80% or so, in light
Learn to show and pray for checking spore germination rate under mirror (40 × 10), 200 spores are at least counted per ware, spore germination rate and suppression is calculated
Rate, standard of sprouting is germ tube length to more than spore minor axis half；
Spore germination rate %=has sprouted spore count/every visual field microscopy spore sum × 100 per the visual field
Suppress spore germination rate %=(control germination rate-processing germination rate)/control germination rate × 100
Linear regression analysis between the niqueMin of relative inhibition and the logarithm value of drug concentration, obtains suppression apple
The valid density (EC50 values) of fruit tree putrefaction disease bacterium spore germination 50%；
The inhibitory action of the Tebuconazole of table 2 and carbendazim and its compounding to Valsa mali spore germination
Note：During SR ＜ 0.5, antagonism is represented；During 0.5≤SR≤1.5, summation action is represented；During SR ＞ 1.5, represent
Synergistic effect
As shown in Table 2, carbendazim, Tebuconazole and its different blended are determined under indoor conditions to rot to apple bark with combination
Germ spore germination shows different inhibitory action effects, it is determined that optimum proportioning；When carbendazim and Original Tebuconazole press 9: 1
Proportioning performance synergistic effect, coefficient of synergism is 6.319, and this proportioning is the optimum proportioning of the two；
The complex preparation of different proportion is obtained to the growth of apple bark rotten pathogenic bacteria silk and spore by carbendazim and Tebuconazole
The inhibitory action of sprouting is different, and it is rotten that the mixture that wherein carbendazim and Tebuconazole are reached with 9: 1 ratio mixtures suppresses apple bark
The coefficient of synergism of rotten germ and spore germination is higher, reaches synergy level.
Influence of 3 paste to the antibacterial band of apple rot pathogen after treatment of different temperature is tested to determine
As agricultural chemicals commodity, shelf life requirements at least (54 ± 2) DEG C storage 14d maintains good emulsus as liniment
Liquid status, simply separate emulsion and water, gently shake that to remain into uniform emulsion qualified, only separate oil reservoir to be unqualified,
And there is not floccule and condensation, good fluidity；Specific method：Appropriate amount sample is taken, is sealed in wide-mouth bottle, (54 ±
2) stored in DEG C insulating box after 14d, see whether oil reservoir and floccule, condensation occur, determine whether product heat storage stability closes
Lattice；
To ensure that the safety of medicament is passed the winter, low tempertaure storage stability test need to be carried out, appropriate amount of sample is fitted into bottle, after sealing
In -15 DEG C of refrigerators, observed after storage 14d, it is not stratified to be qualified；
Using improvement face-off method：The about 30mL PDA culture mediums slightly cooled down after sterilizing are poured into culture dish (d=9cm) to be made
Flat board, is punched with 7mm card punch in flat board center, medicament addition Kong Zhongyu flat boards is maintained an equal level, in hole, surrounding is inoculated with 4 cultures
7mm Valsa mali bacterium disks after 72h, each Jun Panyukong centers are put into 26 DEG C of constant incubator trainings at a distance of about 2.8cm
Support, to add the processing of aqua sterilisa and emulsion paint for control, with 3.315% first sulphur methyl α-naphthyl acetate (smearing formulation) for comparison medicament, often
Processing is repeated 4 times；Each processing bacteriostatic diameter is measured when ware is covered with control and (Fig. 8) is taken a picture；
The fungistatic effect of 3%DW, 5%DW paste is higher than 3.315% under the conditions of normal temperature laboratory (25 ± 2) DEG C as shown in Figure 2
First sulphur methyl α-naphthyl acetate comparison medicament, the significant difference in P=0.05 levels, it can be seen that carbendazim Tebuconazole mixture paste presses down
Bacterium effect is best, therefore selects the experiment of formula progress next step；
As shown in Figure 3,4, fungistatic effect is determined after high temperature (54 ± 2) DEG C, low temperature (- 15 ± 2) DEG C storage 14d, can be with
Find out that difference is not notable under 3%DW paste fungistatic effect and normal temperature after high temperature, low-temperature treatment, 5%DW paste is through high temperature, low temperature
Fungistatic effect illustrates that this formula meets agricultural chemicals with normal temperature phase ratio in 5% horizontal fungistatic effect and significant difference under normal temperature after processing
The shelf life requirements of commodity, and stability is preferable；It is this experiment final products to determine the formula.
Test prevention effect of 4 medicaments to apple tree canker --- isolated shoot is tested
Therapeutic action of the medicament to apple tree isolated shoot
Therapeutic action：1 year green apple tree (Fuji's kind) branch of clip, is cut into 20cm length, is rinsed well with running water, then
Surface sterilization is carried out with 75% alcohol, every card punch with diameter 4mm is punched, and removes bark, the apple tree for cultivating 3d is rotten
Rotten germ bacteria cake (d=4mm) is positioned at branch punching, with preservative film to being wound at punching, bacteria cake is fixed on branch
On, branch two ends are sealed with vaseline；Then branch is put in moisturizing 21h in 26 DEG C of insulating boxs, pathogen is set up with host
Parasitism, then releases moisturizing material, is grouped by the thickness and growing way of branch, the daubing medicament at aperture, not apply medicament
Processing be control, be inserted into damp sand disk, be put in 25 DEG C of tissue culture rooms and carry out moisturizing cultures, after investigation statisticses after control morbidity
The incidence respectively handled, measures Lesion size, calculates lesion area and prevention effect；4 branches of each processing inoculation, each
2 inoculation holes (Fig. 7) of branch；
Prevention effect (%)=[(control lesion area-bacteria cake area)-(processing lesion area-bacteria cake area)]/(control
Lesion area-bacteria cake area) × 100
The incidence of disease=morbidity inoculation points/total inoculation points × 100%
Sick blotchy surface accumulates=1/4 × π × major diameter × minor axis
Therapeutic action of the different agents to isolated shoot apple tree canker the results are shown in Table 4；
Therapeutic action of the different agents of table 4 to isolated shoot apple bark rot
Note：Data are average value ± standard error in table, are represented with same letter after column data through Deng Kenshi duncan's new multiple range methods
Examine not notable in the level differences of P ＜ 0.05, similarly hereinafter.
Table 4 shows that the 3% of the preparation of embodiment 3, extension of the 5%DW paste to scab has certain inhibition；Treatment
5%DW preventive effects highest in experiment, is 93.18%, and secondly 3%DW prevention effects are 92.28%, with the first of comparison medicament 3.315%
Sulphur methyl α-naphthyl acetate difference is not notable, and wherein 5%DW prevention effects are significantly higher than 3.315% first sulphur methyl α-naphthyl acetate；
Test the prevention effect that 5 different agents are rooted out to apple bark rot scab
Valsa mali is inoculated with tree body trunk in May, 2014, after after its morbidity, the typical scab of selection uses Fan Shi
The thorough scraped clean of diseased tissues is around exposed health tissues and scrapes 0.5cm or so again outward by scraper；Wound is smeared using medicament
And surrounding health tissue, the processing of setting：3%DW paste, 5%DW paste, 3.315% first sulphur methyl α-naphthyl acetate are comparison medicament, breast
Shellac varnish control, sterilized water are blank control；5 pieces of scabs are often handled, are repeated 3 times；
Inoculation method：Select side shoot more and the smooth disease-free apple tree of trunk, with 4mm card punch under trunk side shoot
The smooth place punching at end, an equal amount of sick bark tissue is filled into handled hole, with the absorbent cotton for being moistened with sterilized water
Moisturizing, then around trunk absorbent cotton is snarled and obturaged with preservative film；Preservative film and absorbent cotton are peeled off after processing tree morbidity, observation
Incidence；
Strike off and handled after scab on May 20th, 2014, after the recurrence feelings that scab is investigated, recorded after control recurrence
Condition, the width of vernier caliper measurement callus, and calculate prevention effect；As shown in Figure 4；
Prevention effect (%)=(control recurrence rate-processing recurrence rate)/control recurrence rate × 100
Influence of the different disposal of table 6 to the eradicant action of apple bark rot scab and to apple tree Callus formation
Strike off and 3%, 5%DW paste, the first sulphur methyl α-naphthyl acetate of comparison medicament 3.315%, auxiliary agent emulsion paint 120 days are smeared after scab
After investigate each 15 pieces of scab recurrence of processing, blank control has 8 pieces of recurrences, and emulsion paint processing recurs 2 pieces, and preventive effect is
13.33% (being shown in Table 6)；3%th, the callus width that is formed is 9.07mm, 8.53mm after the processing of 5%DW paste, between the two
Difference is not notable, with CK processing compare to form significant difference, with comparison medicament first sulphur methyl α-naphthyl acetate without significant difference.
Finally by Integrated comparative, it is this test products that 3%DW paste is determined for the time being.
The liniment of embodiment 1 (paste)
Carbendazim：2.7%th, Tebuconazole：0.3%th, Emulsifier O p10：10.20%~10.70%, gibberellin GA：
0.002%th, alcohol (4.3 ± 0.5) % emulsion paints polishing is to 100g
Each composition of above-mentioned formula is accurately weighed first, and carbendazim and Tebuconazole are put into agitator tank, plus appropriate emulsifying agent
Op10 is sufficiently stirred for after dissolving, then is stirred with being put into after small beaker precise gibberellin, plus suitable alcohols dissolving in agitator tank
Uniformly, emulsion paint polishing is to 100g.
The liniment of embodiment 2 (paste)
Carbendazim：4.5%th, Tebuconazole：0.5%th, Emulsifier O p10：15.94%~16.50%, gibberellin GA：
0.002%th, alcohol (4.3 ± 0.5) % emulsion paints polishing is to 100g.Preparation process is shown in embodiment 1.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
Any modifications, equivalent substitutions and improvements made within refreshing and principle etc., should be included in the scope of the protection.
Claims (3)
1. a kind of composite bactericide, it is characterised in that the composite bactericide includes according to parts by weight：Carbendazim 2.7g, penta azoles
Alcohol 0.3g, 0p10：10.20~10.70g, gibberellin GA：0.002g, 4.3 ± 0.5g of alcohol, emulsion paint polishing to 100g；
Carbendazim, Tebuconazole are respectively active compound；
Composite bactericide is liniment.
2. a kind of composite bactericide as claimed in claim 1, it is characterised in that the composite bactericide preparation method includes following
Step：
Take 2.7g carbendazim, 0.3g Tebuconazoles to be put into agitator tank, plus 10.20~10.70g Emulsifier Os p10 be sufficiently stirred for dissolving
Afterwards, stand well mixed, then weigh gibberellin 0.002g with small beaker, be put into after ± 0.5g alcohol dissolving of plus 4.3 in agitator tank
Stir, emulsion paint polishing to 100g.
3. composite bactericide as claimed in claim 1, it is characterised in that the composite bactericide application method, is applied using limb
Smear, sick scar scraped clean is smeared at trunk.
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