CN110199990B - Application of surfactant as auxiliary agent in aspect of improving effective utilization rate of fruit and vegetable pesticide - Google Patents

Application of surfactant as auxiliary agent in aspect of improving effective utilization rate of fruit and vegetable pesticide Download PDF

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CN110199990B
CN110199990B CN201910519159.7A CN201910519159A CN110199990B CN 110199990 B CN110199990 B CN 110199990B CN 201910519159 A CN201910519159 A CN 201910519159A CN 110199990 B CN110199990 B CN 110199990B
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isaria fumosorosea
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王宏民
赵义涛
张仙红
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Shanxi Agricultural University
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N25/00—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests
    • A01N25/30—Biocides, pest repellants or attractants, or plant growth regulators, characterised by their forms, or by their non-active ingredients or by their methods of application, e.g. seed treatment or sequential application; Substances for reducing the noxious effect of the active ingredients to organisms other than pests characterised by the surfactants
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
    • A01N63/30—Microbial fungi; Substances produced thereby or obtained therefrom

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Abstract

Disclosure of the inventionThe surfactant is used as an auxiliary agent for improving the effective utilization rate of the fruit and vegetable pesticide; the surfactant refers to PEG-12 polydimethylsiloxane (OFX-0193) and sodium diisobutylnaphthalenesulfonate (Nekal); the biological insecticide refers to: isaria fumosorosea PF 904; the fruit and vegetable means that: cabbage, apple, eggplant, and cucumber. The results show that: 2 surfactants in 125 mg.L‑1‑500 mg·L‑1The wetting performance of the isaria fumosorosea spore suspension on the leaves of 4 test plants can be improved. The measurement result of the spore germination inhibition rate shows that: nekal and OFX-0193 have no inhibition effect on spore germination of Isaria fumosorosea PF904, so that Nekal and OFX-01932 surfactants can be used as an auxiliary agent for controlling apple and cabbage pests by Isaria fumosorosea; OFX-0193 can also be used as an auxiliary agent for controlling solanaceous pests.

Description

Application of surfactant as auxiliary agent in aspect of improving effective utilization rate of fruit and vegetable pesticide
Technical Field
The invention belongs to the technical field of pesticide adjuvant application. In particular to application of a surfactant as an auxiliary agent in improving the effective utilization rate of fruit and vegetable pesticides.
Background
The wettability and the retention of the liquid medicine on a target plant directly influence the effective utilization rate of the pesticide. The wettability is related to the surface tension of the liquid medicine, the contact angle of the liquid medicine on the blade, and the like. The surface active agent is added, so that the surface tension of the pesticide liquid can be effectively reduced, the contact angle between the pesticide liquid and the target can be reduced, the wetting capacity and the deposition of the pesticide on the target plant can be effectively improved, the loss of the pesticide can be reduced, and the pollution of the pesticide to the environment can be reduced. However, the application technology of domestic pesticides is relatively laggard, most of the surfactants added in the liquid medicine do not reach the critical micelle concentration, so that the utilization rate of the pesticide is low, and particularly, no research report of adding the surfactants exists so far. In addition, as the wetting and retention performances of the liquid medicine can be influenced by the biological characteristics of the microstructures and the like of different plant leaves, the screening of the surfactant types suitable for the application of the biological pesticide and different plants has important significance for improving the effective utilization rate of the biological pesticide, reducing the dosage and saving the cost. A great deal of research shows that a lot of liquid medicines are difficult to adhere to the surfaces of plants such as cabbage, apple and the like and roll off, so that the pollution of pesticide to the environment is caused, and therefore, the improvement of the physicochemical property of the liquid medicine for pesticide application, particularly the reduction of the surface tension of the liquid medicine is an important way for improving the utilization rate of biological pesticide. In recent years, with the increasing use of biological pesticides year by year, screening of appropriate auxiliary agents has important significance for improving the control effect of the biological pesticides.
The invention relates to different surfactants comprising: the water-soluble silicone oil OFX-0193 (Dow Corning Shanghai Co., Ltd.) is mainly used as a cosmetic additive or a surfactant, is very stable in a water system, and can reduce the surface tension; the sodium diisobutylnaphthalenesulfonate Nekal is mainly used for penetrants, emulsifiers, detergents, dyeing assistants, dispersants, wetting agents and the like.
Isaria fumosorosea is an entomopathogenic fungus with great development potential, and has important pathogenic effects on vegetable pests such as Plutella xylostella (Linnaeus), Bemisa babaci (Gennadius), Myzus persicae (Sulzer) and the like, and fruit tree pests and tea tree pests. Whether spores of entomopathogenic fungi can attach to the target plant is critical for its infection of the host insect. However, most of these spores lack wettability, dispersibility and systemic property, so that the control effect of microbial pesticides on pests can be better exerted only by attaching entomopathogenic fungal spores to target plants as much as possible. At present, many studies are made on the use of surfactants for improving the utilization efficiency of chemical pesticides, and no report is made as to whether the surfactants can improve the adhesion of biological pesticides to host plants and the efficacy of the biological pesticides.
Disclosure of Invention
Therefore, the Isaria fumosorosea spore suspension added with different surfactants is used as a test material, the surface tension of the liquid medicine, the contact angle and the maximum stable retention amount on different test plant leaves are measured, and the safety evaluation of the test surfactant is carried out, so as to screen out the optimal surfactant and concentration for improving the effective utilization rate of Isaria fumosorosea PF 904. The surfactant is used as an auxiliary agent for improving the effective utilization rate of the biological insecticide, and the report on the effective utilization rate of the biological insecticide at home and abroad is not available at present.
In order to achieve the purpose, the invention discloses the following technical contents:
the surfactant is used for improving the application of the biological insecticide in the aspect of effective utilization rate of fruit and vegetable pests; the surfactant auxiliary agent refers to PEG-12 polydimethylsiloxane (OFX-0193) and sodium diisobutylnaphthalenesulfonate (Nekal); the pesticide refers to: isaria fumosorosea PF 904; the fruit and vegetable means that: cabbage, apple, eggplant, and cucumber.
The effective utilization rate of the invention refers to that the surfactant can improve the wetting performance of Isaria fumosorosea PF904 on fruit and vegetable plants, reduce the contact angle and the maximum stable retention amount of liquid medicine.
For screening Isaria fumosorosea (Isaria fumosorosea) PF904, the wetting properties, maximum retention and contact angle of Isaria fumosorosea on fruit and vegetable leaves after adding different surfactants were studied in this experiment. The results show that: PEG-12 polydimethylsiloxane (OFX-0193), alpha-sodium alkenyl sulfonate (AOS) and sodium diisobutylnaphthalenesulfonate (Nekal) 3 surfactants at 125 mg.L-1-500 mg·L-1The wettability of the isaria fumosorosea spore suspension on the leaves of 4 test plants can be improved; wherein the concentration is 250 mg.L-1Maximum retention of Nekal on cabbage and apple leaves (11.90, 13.28) mg cm-2The contact angle (29.5 degrees and 27.5 degrees) is obviously better than that of OFX-0193 and AOS, and the wetting effect is best; adding 125 mg.L onto eggplant leaf-1OFX-0193 (maximum retention capacity of 9.45 mg/cm)-2(ii) a And the solution surface tension when the Pullulan (Pullulan) reaches the critical micelle concentration is greater than the critical surface tension of cabbage, apple and eggplant leaves, the contact angles are all greater than 90 degrees, and the wetting property is the worst. The measurement result of the spore germination inhibition rate shows that: nekal and OFX-0193 have no inhibitory effect on the germination of Isaria fumosorosea PF904 spore, and AOS has an inhibitory effect on the spore. Therefore, the surfactants Nekal and OFX-01932 can be used as an auxiliary agent for controlling apple and cabbage pests by using isaria fumosorosea; OFX-0193 can also be used as an auxiliary agent for controlling solanaceous pests.
The invention mainly solves the problem that the biological insecticide isaria fumosorosea is not easy to attach to the surface of a plant to seriously affect the pesticide effect, mainly inspects the influence of different surfactants on the wetting performance of liquid medicine and the safety of the liquid medicine to the biological insecticide, and has the main difficulty in the research of 2 surfactants on the aspects of reducing the surface tension and the contact angle of the liquid medicine and improving the stable retention amount of the liquid medicine.
Drawings
FIG. 1 is a graph showing the surface tension corresponding to the critical micelle concentration reached by 4 adjuvants;
FIG. 2 shows the results of the contact angles of Isaria fumosorosea spore suspensions on leaves of test plants with different surfactants added; wherein A: cabbage; b: apple, apple; c: eggplant; d: cucumber. Data in the figure are mean ± standard deviation. Different letter designations were significantly different at a P <0.05 level as tested by the LSD method.
FIG. 3 is the maximum stable retention of Isaria fumosorosea spore suspensions on cabbage and apple leaves with different surfactants added; a: cabbage; b: an apple. Data in the figure are mean ± standard deviation. The different letter representations are examined by the LSD methodP<0.05The level difference was significant.
FIG. 4 is the maximum stable retention of Isaria fumosorosea spore suspensions on eggplant and cucumber leaves with different surfactants added; wherein A: eggplant; b: cucumber. Data in the figure are mean ± standard deviation. Different letter designations were significantly different at a P <0.05 level as tested by the LSD method.
FIG. 5 shows the germination of Isaria fumosorosea conidia after the addition of different surfactants; wherein A, E: spore germination in control; B. f: spore germination in OFX-0193; C. g: spore germination in Nekal; D. h: spores germinate in AOS. A-D: 12h of conidium germination; E-H: conidiophores germination condition 24 h. CO: conidia; GT: a germ tube; AP: and (5) attaching spores.
Detailed Description
The invention is described below by means of specific embodiments. Unless otherwise specified, the technical means used in the present invention are well known to those skilled in the art. In addition, the embodiments should be considered illustrative, and not restrictive, of the scope of the invention, which is defined solely by the claims. It will be apparent to those skilled in the art that various changes or modifications in the components and amounts of the materials used in these embodiments can be made without departing from the spirit and scope of the invention. The raw materials and reagents used in the present invention are commercially available.
Example 1
1 materials and methods
1.1 materials
Leaf of the test plant: fresh leaves of cabbage, apple, eggplant and cucumber planted in experimental field of Shanxi university of agriculture are collected.
Test strains and surfactants: isaria fumosorosea PF904 (stored in the insect emphasis laboratory of Shanxi university of agriculture, see the bioengineering bulletin, 2019,35 (1): 114-; PEG-12 polydimethylsiloxane (OFX-0193) (Dow Corning Shanghai Co., Ltd.), alpha-sodium alkenyl sulfonate AOS (Linyi city Lvson chemical Co., Ltd.), sodium diisobutylnaphthalenesulfonate Nekal (Linyi city Lvson chemical Co., Ltd.), and Pullulan (Shandong Corna Biotech Co., Ltd.).
The test instrument: JC2000C1 contact angle measuring instrument (Shanghai Zhongchen digital technology equipment Co., Ltd.), JYW-200B micro-control surface automatic tensiometer (Chengdui and instruments manufacturing Co., Ltd.), electronic balance (Beijing Sidoolis scientific instruments Co., Ltd.), 1-100 mu L of liquid-transferring gun (Shanghai Kenqing machine Co., Ltd.), and the like.
1.2 methods
1.2.1 measurement of the critical surface tension of leaves of test plants
Selecting fresh cabbage, apple, eggplant and cucumber leaves, dripping liquid with different surface tensions on the leaves, and measuring a contact angle by using a contact angle measuring instrument. And (3) making a regression equation of the contact angle cos theta to the surface tension value of the solution, wherein when cos theta =1, the corresponding liquid surface tension is the critical surface tension value of the plant.
1.2.2 preparation of spore suspensions
Activating culture medium of Isaria fumosorosea PF904 strain stored at 4 deg.C at room temperature for 1d, inoculating on PDA culture medium, culturing for 10d, slowly scraping off spore with inoculating needle, repeatedly washing with clear water to obtain spore suspension, dripping the spore suspension on hemocytometer with pipette gun,adjusting spore concentration to 1 × 104spores/mL.
1.2.3 preparation of test solutions
Adding different surfactants into spore suspension prepared from Isaria fumosorosea, respectively preparing into spore suspension containing surfactant 62.5 mg.L-1、125 mg·L-1、250 mg·L-1、500 mg·L-1And 1000 mg. L-15 liquid medicines with gradient mass concentration, and spore suspension added with clear water is used as a control.
1.2.4 measurement of surface tension and Critical micelle concentration
And measuring the surface tension of the liquid medicine with corresponding concentration by using a micro-control meter interface automatic tensiometer. The test solution is poured into a 250mL triangular flask respectively, the flask is shaken fully and uniformly, the solution to be tested is poured into a glass vessel and placed on an automatic lifting table, the glass vessel is clicked to ascend until a platinum ring is completely immersed into the solution, the glass vessel is clicked to descend until the platinum ring is completely pulled open the liquid level, and an interfacial tension meter can record that the maximum value of the pulling force is the surface tension of the solution. The same sample is repeatedly measured for 3 times, the measured surface tension and the concentration are plotted to obtain a curve, and the inflection point of the curve is the critical micelle concentration of the surfactant in the liquid medicine.
1.2.5 determination of contact Angle
Cut 2X 2cm2Fresh and clean plant leaves for test (avoiding veins, scabs and the like), adhering the plant leaves on a glass slide by using a double-sided adhesive tape, horizontally placing the plant leaves on a sample platform of a contact angle measuring instrument, sucking prepared liquid medicine by using a liquid transfer gun and dripping the liquid medicine on the leaf surfaces, taking a camera on the contact angle measuring instrument to take a photograph of liquid drops on the leaf surfaces at the time of 20s, inputting the liquid drops into a computer, calculating the contact angles of the liquid drops on different plant leaves by a quantity angle method, taking isaria fumosorosea suspension without adding an auxiliary agent as a reference, and controlling the temperature to be (25 +/-1) DEG C during measurement.
1.2.6 determination of maximum Stable Retention
The cutting area is 2 multiplied by 2cm2The 4 kinds of leaves of the plant to be tested are weighed by balance (W0, g), vertically put into the liquid medicine to be tested for 3-5 s by being held by tweezers, quickly pulled out of the water surface, vertically suspended until no more liquid drop flow existsWeighing (W1, g) while flowing, and calculating the maximum stable retention R of the bladem(mg·cm-2). Maximum stable retention Rm(mg·cm-2) = (W1-W0) × 1000/blade area (cm)2)。
1.2.7 Effect of surfactants on spore germination
Collecting Pf904 conidia of Isaria fumosorosea cultured on PDA plate for 10 days, adding into sterile water containing 2% glucose, oscillating uniformly on vortex oscillator, and adjusting spore concentration to 10 with blood counting chamber4spores/mL. Adding the surfactant which is screened by the method of 1.2.4-1.2.6 and can improve the wettability of Isaria fumosorosea to the spore suspension at the optimal concentration. Spore suspensions containing different types and concentrations of surfactants were cultured on a shaker at 25 + -1 deg.C and 160r/min with shaking, and each concentration was repeated 5 times, using spore suspension without surfactant as control. And (5) dripping 20 mu L of spore suspension on a concave glass slide at 12h and 24h respectively, placing the concave glass slide under a microscope for counting the spore germination condition, and photographing and recording. Spore germination inhibition (%) = (1-number of treated sprouts/number of control sprouts) × 100%.
1.3 data analysis
The test data are calculated by Microsoft Excel 2010, the graph is plotted by Sigmaplot 12.5, the statistical analysis is carried out by using DPS 7.05 software, the LSD method is used for testing the difference significance, and the significance level is set asp＜0.05。
2 results and analysis
2.1 Critical surface tension of leaves of the plant to be tested
The test measures the critical surface tension values of cabbage, apple, eggplant and cucumber leaves (table 1). When the surface tension of the liquid medicine is less than the critical surface tension of the plant, the liquid medicine can be wetted and spread on the plant leaves.
TABLE 14 regression equation and critical surface tension of plants
Figure 858677DEST_PATH_IMAGE001
2.2 surface tension of different test solutions
The surface tension value of the isaria fumosorosea spore suspension is 70.8mN/m, which is obviously higher than the critical surface tension of leaves of cabbage, apple, eggplant and cucumber of the test plants, and the leaves are difficult to wet and spread on the leaves of the test plants. When the added concentrations of Pullulan, OFX-0193, AOS and Nekal reach 500 mg.L respectively-1、500 mg·L-1、250 mg·L-1And 250 mg. L-1The surface tension of the liquid medicine was reduced to 45.6 mN · m-1、27.3 mN·m-1、24.3 mN·m-1And 21.8 mN · m-1And the surface tension values of the 4 adjuvant solutions no longer changed with increasing concentration, i.e. the Critical Micelle Concentration (CMC) was reached.
From the measured surface tensions (fig. 1) corresponding to the 4 auxiliary agents reaching the critical micelle concentration, when OFX-0193, AOS and Nekal 3 auxiliary agents are added, the critical surface tensions of the 4 tested leaves are all larger than the surface tensions thereof, which indicates that the 3 auxiliary agents can significantly improve the wetting performance of the liquid medicine under the appropriate concentration, wherein Nekal is the most preferable, and the surface tensions corresponding to Pullulan reaching the critical micelle concentration are always larger than the critical surface tensions of cabbage, apple and eggplant leaves.
2.3 contact angles of liquid medicines added with different surfactants on 4 plant leaves
The contact angle results (FIG. 2) of Isaria fumosorosea spore suspensions with different surfactants on leaves of test plants show that: with the increase of the OFX-0193, AOS and Nekal concentrations, the contact angle of the isaria fumosorosea spore suspension on each test plant leaf is obviously reduced, and the surfactant at each concentration is obviously different from that of a control group. On cabbage leaves, when the concentration of Nekal and AOS is only 62.5 mg.L-1When the contact angle is less than 60 degrees, the wettability is good, and the contact angles of 5 concentrations of OFX-0193 and Pullulan are both more than 60 degrees, and the wettability is medium to poor; on apple leaves, when the concentration of Nekal, OFX-0193 and AOS is only 62.5 mg.L-1When the contact angles are less than 60 degrees, the wettability is good, the contact angles of 5 concentrations of Pullulan are between 90 and 100 degrees, and the wettability is poor; adding Nekal, OFX-0193 and AOS to eggplant and cucumber leaves at a concentration of 62.5 mg.L-1When the contact angle is less than 60 degrees, the wettability is good, and the added concentration of Pullulan is 250 mg.L-1、500 mg·L-1When the contact angle between cucumber leaf and eggplant leaf is less than 60 deg., its wettability is good. Considering that the concentration of the surfactant in the liquid medicine should be as low as possible, and the auxiliary agent with high surface tension reducing efficiency needs to be selected, OFX-0193, AOS and Nekal are selected for the next test.
2.4 maximum stable retention of medicinal liquid added with different surfactants on 4 plant leaves
2.4.1 maximum Stable Retention of test solution on cabbage and apple leaves
Retention of isaria fumosorosea spore suspension on cabbage and apple leaves after addition of surfactant (figure 3) shows: the liquid holdup of Isaria fumosorosea spore suspension without adding surfactant in cabbage and apple leaves is only 3.79 mg cm-2、4.17 mg·cm-2The retention capacity rises significantly with increasing concentrations of the 3 auxiliaries. When the concentration of Nekal and AOS is 250 mg.L-1The concentration of OFX-0193 is 500 mg.L-1(critical micelle concentration is reached), the retention of the test solution on the cabbage and apple leaves is maximized. Wherein 250 mg.L is added-1The maximum retention of the Nekal liquid medicine on the leaves of 2 test plants can reach 11.90 mg cm-2、13.28 mg·cm-2Both significantly higher than the other 2 surfactants and the control.
2.4.2 maximum Stable Retention of test solution on eggplant and cucumber leaves
The maximum stable retention of isaria fumosorosea spore suspension on eggplant and cucumber leaves after addition of surfactant (fig. 4) shows: the liquid holdup of Isaria fumosorosea spore suspension without adding surfactant in eggplant and cucumber leaf is more, respectively 8.52 mg cm-2、7.66 mg·cm-2. With the increasing concentration of the auxiliary agent, the retention of the test solution on eggplant and cucumber leaves is reduced, the retention of cabbage and apple leaves is OFX-0193 at the minimum, and the retention of eggplant and cucumber leaves is the maximum. When the added concentration of OFX-0193 is 125 mg.L-1In the meantime, of the liquid medicineThe surface tension reaches the critical surface tension of eggplant, and the retention of OFX-0193 liquid medicine on eggplant leaves reaches 9.45 mg cm at most-2。
2.5 Effect of the test Agents on Isaria fumosorosea PF904 Germination
The inhibition of isaria fumosorosea spore germination by the optimal wetting concentration of 3 surfactants is shown in table 2. The effect of different test surfactants on Isaria fumosorosea conidiophore germination was different. Wherein OFX-0193 and Nekal have no inhibition effect on the germination of Isaria fumosorosea spores (figure 5B, F, C, G), and after the Isaria fumosorosea conidia added with OFX-0193 is cultured for 24h, a large number of attached spores can be observed (figure 5F), part of spores can germinate towards two ends, which shows that OFX-0193 has a certain promotion effect on the germination of Isaria fumosorosea spores; the AOS had a clear effect on the germination of Isaria fumosorosea spores (FIG. 5D, H), when the concentration was 125 mg. L-1When the spore germination inhibition rate is 19.58%, the optimal wetting concentration is 250 mg.L-1The spore germination inhibition rate reaches 85.71%. It can be seen that Nekal and OFX-0193 can be used as surfactants for Isaria fumosorosea PF904, whereas AOS cannot be used as a surfactant for Isaria fumosorosea PF904 due to inhibition of spore germination.
TABLE 2 inhibition of Isaria fumosorosea PF904 spore germination by different surfactants
Figure 720322DEST_PATH_IMAGE002
Note: treating for 24h to obtain spore germination inhibition rate; the different letter representations are examined by the LSD methodP<0.05The level difference is significant
And (4) conclusion: PEG-12 polydimethylsiloxane (OFX-0193) and sodium diisobutylnaphthalenesulfonate (Nekal) 2 surfactants at 125 mg.L-1-500 mg·L-1The wetting performance of the isaria fumosorosea spore suspension on leaves of 4 plants including cabbage, apple, eggplant and cucumber can be improved, the stable retention amount and the contact angle are improved, and the germination of the isaria fumosorosea spores is not influenced, so that the 2 surface activitiesThe preparation can be used as an effective assistant of biopesticide isaria fumosorosea. The Pullulan (Pullulan) has poor wetting performance, and alpha-sodium alkenyl sulfonate (AOS) has obvious inhibition effect on Isaria fumosorosea spore germination and cannot be used as an effective auxiliary agent of Isaria fumosorosea.

Claims (2)

1. The application of a surfactant PEG-12 polydimethylsiloxane (OFX-0193) as an auxiliary agent for improving the wettability of a biopesticide Isaria fumosorosea PF904 on apple leaves.
2. The use as claimed in claim 1, wherein the wetting property is that the surfactant OFX-0193 lowers the contact angle of the liquid medicine with the hydrophobic apples leaves and increases the stable retention of the agent on the leaves.
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