CN113575235A - Method for synergistically preventing and treating Tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments - Google Patents
Method for synergistically preventing and treating Tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments Download PDFInfo
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Abstract
The invention provides a method for synergistically preventing and controlling tetranychus urticae by using small Amblyseius barkeri and a pyrethroid medicament, wherein the small Amblyseius barkeri is the small Amblyseius barkeri with pyrethroid resistance. The invention connects small neoseiulus barkeri with pyrethroid drug resistance to the vegetable seedlings with two-spotted spider mite, then sprays pyrethroid pesticide on the leaf surfaces of the vegetable seedlings, and carries out field management. The invention can effectively solve the contradiction between biological control and chemical control in the control process of the tetranychus urticae koch, and the application of the chemical agent can be effectively reduced by using the drug-resistant neoseiulus barkeri combined with the chemical agent to control the tetranychus urticae koch, thereby ensuring the ecological safety and the human health and being a very good comprehensive measure for controlling pests.
Description
Technical Field
The invention relates to the field of biological control of agricultural and forestry pests, and in particular relates to a method for synergistically controlling tetranychus urticae by small neoseiulus barkeri and a pyrethroid medicament.
Background
The Neoseiulus pasteurianus bushri belongs to the genus Neoseiulus of Phytoseiidae, is an important natural enemy of thrips and spider mites, and is widely applied in Australia, Europe and other countries. The mite has the characteristic of polyphagia, can also take scale insects, aphids, psyllids, nematodes and the like as food, and is called one of the most effective natural enemies for biological control because of the advantages of strong diffusion, low mortality, short development period, easy feeding, mass propagation and the like.
Biological control and chemical control are effective measures widely adopted in the comprehensive treatment of agricultural pests, and especially, chemical agent control plays an important role in controlling pests. However, the long-term use of chemical pesticides often causes environmental pollution and the increase of drug resistance of pests, and even leads to the death of a large number of natural enemies of pests. Therefore, the natural enemies with drug resistance are applied to field release for controlling pests, so that the pests with drug resistance can be effectively controlled, and the death of the natural enemies can be avoided. On the premise that chemical prevention and control are still the main factors for pest control at present, drug resistance screening and application of phytoseiid mites are particularly important.
Disclosure of Invention
The invention aims to provide a method for preventing and treating Tetranychus urticae by synergy of neoseiulus barkeri and pyrethroid medicaments, which can effectively solve the contradiction between biological prevention and chemical prevention and treatment in the process of preventing and treating Tetranychus urticae.
The technical scheme adopted by the invention is as follows:
a method for synergistically controlling Tetranychus urticae with a pyrethroid medicament, wherein the Neoseiidae is pyrethroid-resistant.
The resistance of the neoseiulus barkeri to the deltamethrin is incomplete dominant polygenic inheritance.
Preferably, the pyrethroid medicament is cyhalothrin, deltamethrin, bifenthrin, lambda-cyhalothrin, ethofenprox, beta-cypermethrin, cypermethrin; the acaricide is etoxazole; the organic phosphorus medicament is chlorpyrifos; any one of the second generation new nicotinoyl insecticides thiamethoxam.
Preferably, the small neoseiulus barkeri with pyrethroid resistance is grafted to the vegetable seedlings with two-spotted spider mites, and then the pyrethroid pesticide is sprayed on the leaf surfaces of the vegetable seedlings for field management.
Preferably, the small neoseiulus barkeri with pyrethroid resistance is grafted to the vegetable seedlings with tetranychus urticae according to the benefit-to-pest ratio of 1:5-20, and cyhalothrin or deltamethrin with the concentration of 5 x 10 < -3 > to 10 x 10 < -3 > g/L is sprayed on the leaf surfaces of the vegetable seedlings for field management.
Preferably, the small neoseiulus barkeri with pyrethroid resistance is connected to the vegetable seedlings with tetranychus urticae according to the benefit-to-pest ratio of 1:10, and then 2.5 percent cyhalothrin emulsifiable concentrate or 3000 times of 2.5 percent deltamethrin emulsifiable concentrate diluent is sprayed on the leaf surfaces of the vegetable seedlings for field management; the application concentration of cyhalothrin or deltamethrin is 8.3 multiplied by 10-3 g/L.
Preferably, the breeding method of the small neoseiulus barkeri with pyrethroid resistance comprises the following steps:
picking out female adult mite of Amblyseius barkeri from parent, and applying 98% deltamethrin crude drug to LC of female adult mite25The mites are treated by dosage, and after 24 hours, the surviving neoseiulus barkeri is transplanted into the pink mites; meanwhile, in the process of feeding the flour mites, deltamethrin agents with the same concentration are sprayed in the feed, the agent spraying concentration is gradually increased along with the increase of drug resistance of flour mite populations, the new small amblyseius barkeri is fed by the drug-resistant flour mites, and the steps are repeated and screened for multiple generations.
Preferably, LC is selected25Dosage of 0.14mg/L as BabbittThe initial concentration of the neoseiulus resistance breeding is that under the breeding of the deltamethrin pesticide with the initial concentration, the mortality of the neoseiulus pasteurianus in 24 hours is 79.01-61.22%, each generation of the subsequent screening is carried out, the pesticide concentration controls the mortality of the neoseiulus pasteurianus to be 52.11-86.32%, and the neoseiulus pasteurianus with pyrethroid resistance is bred for 30 generations.
Preferably, the method for breeding the small neoseiulus barkeri with pyrethroid resistance is as follows:
placing sponge (diameter 4.5cm and thickness 0.5cm) and absorbent cotton (diameter 4.0cm) in a culture dish with diameter 5cm from bottom to top, placing in a stainless steel tray (50cm × 35cm), and adding distilled water to soak the sponge and absorbent cotton to form a floating shape;
cutting cowpea leaves damaged by Tetranychus urticae, making into a leaf disc with diameter of 2.5cm, keeping the density of Tetranychus urticae at 40-60 heads/disc, supplementing when the number is insufficient, and placing the back of the leaf upwards on absorbent cotton;
respectively picking eggs laid by the Amblyseius barkeri for less than 6h, placing single eggs into the prepared leaf disc, and feeding the eggs in a single way under the environmental conditions of room temperature of 25 ℃, RH 80-90% and photoperiod of 14L: 10D.
The invention also provides small neoseiulus barkeri with pyrethroid resistance, which is cultivated by the breeding and reproduction method.
The invention has the advantages that:
1. the invention can effectively solve the contradiction between biological control and chemical control in the control process of the tetranychus urticae koch, and the application of the chemical agent can be effectively reduced by using the drug-resistant neoseiulus barkeri combined with the chemical agent to control the tetranychus urticae koch, thereby ensuring the ecological safety and the human health and being a very good comprehensive measure for controlling pests.
2. The deltamethrin or cyhalothrin and the neoseiulus barkeri drug-resistant strain are combined to prevent and treat tetranychus urticae so as to achieve better prevention and treatment effect; compared with the single use of deltamethrin or cyhalothrin, the combined use of the new small neoseiulus drug-resistant strain with deltamethrin or cyhalothrin has the advantages that the control period (14d) is shortened by 57 percent, the loss of economic crops can be greatly reduced by shortening the control period, and the economic effect is obvious.
3. The deltamethrin or cyhalothrin and the neoseiulus barkeri drug-resistant strain are combined to prevent and treat tetranychus urticae so as to achieve better prevention and treatment effect; after the 14 th day of the combined application of the deltamethrin or cyhalothrin new small amblyseius drug-resistant strain, the control effect on tetranychus urticae is 100%, and the number of the insect population is not increased until the 22 th day of control. However, the application of two pyrethroid medicaments alone and the combined application of the two medicaments and the neoseiulus barkeri sensitive strain have the tendency that the population number of the population of. The new Amblyseius barkeri drug-resistant strain and the pyrethroid pesticide are jointly applied, so that the control efficiency can be improved, the control time of the Tetranychus urticae is prolonged, and manpower and material resources are saved.
4. The resistance of the neoseiulus pasteurianus to the deltamethrin generated by the method belongs to incomplete dominant polygenic inheritance, and the genetic mode can lead the resistance population of the neoseiulus pasteurianus to develop rapidly; the application of the drug-resistant predatory mites in the field can reduce the contradiction between biological control and chemical control, and create favorable conditions for comprehensively controlling pests.
5. After 30 generations of breeding, the concentration of the breeding agent is finally increased to 30mg/L, and the deltamethrin is used for LC of the Amblyseius barkeri50The value is improved from 0.32mg/L to 72.44mg/L, and the drug resistance multiple is 226.38 times.
Drawings
FIG. 1 Amblyseius barkeri parent SS, anti-deltamethrin RR and hybrid F1LC-P lines for SR and RS;
FIG. 2 depicts the resistant, sensitive and backcross progeny BC of neoseiulus pasteurii1(F1RSMale parent multiplied by RR male parent) actual dose and BC-E1A desired dose response curve;
FIG. 3 depicts the resistant, sensitive and backcross progeny BC of Amblyseius barkeri1(F1SRMale parent multiplied by SS male parent) actual dose and BC-E2A dose response curve is expected.
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention. The experimental procedures in the following examples are conventional unless otherwise specified. Materials, reagents, instruments and the like used in the following examples are commercially available unless otherwise specified.
Example 1
The breeding method of the small neoseiulus barkeri (abbreviated as small neoseiulus barkeri sensitive line) with pyrethroid resistance comprises the following steps:
picking out female adult mite of Amblyseius barkeri from parent, and applying 98% deltamethrin crude drug to LC of female adult mite25The mites are treated by dosage, and after 24 hours, the surviving neoseiulus barkeri is transplanted into the pink mites; meanwhile, in the process of feeding the flour mites, deltamethrin agents with the same concentration are sprayed in the feed, the agent spraying concentration is gradually increased along with the increase of drug resistance of flour mite populations, the new small amblyseius barkeri is fed by the drug-resistant flour mites, and the steps are repeated and screened for multiple generations.
In this embodiment, LC is selected25The dose of 0.14mg/L is used as the initial concentration of the resistance breeding of the small neoseiulus barkeri, under the breeding of the deltamethrin medicament with the concentration, the mortality of the small neoseiulus barkeri in 24 hours is between 79.01 and 61.22 percent, each generation of screening is carried out, the concentration of the medicament is used to control the mortality of the small neoseiulus barkeri to be between 52.11 and 86.32 percent, and the small neoseiulus barkeri with pyrethroid resistance is obtained by breeding for 30 generations.
Example 2
The breeding method of the small neoseiulus barkeri (abbreviated as) with pyrethroid resistance comprises the following steps:
placing sponge (diameter 4.5cm and thickness 0.5cm) and absorbent cotton (diameter 4.0cm) in a culture dish with diameter 5cm from bottom to top, placing in a stainless steel tray (50cm × 35cm), and adding distilled water to soak the sponge and absorbent cotton to form a floating shape;
cutting cowpea leaves damaged by Tetranychus urticae, making into a leaf disc with diameter of 2.5cm, keeping the density of Tetranychus urticae at 40-60 heads/disc, supplementing when the number is insufficient, and placing the back of the leaf upwards on absorbent cotton;
respectively picking eggs laid by the Amblyseius barkeri for less than 6h, placing single eggs into the prepared leaf disc, and feeding the eggs in a single way under the environmental conditions of room temperature of 25 ℃, RH 80-90% and photoperiod of 14L: 10D.
Example 3
A method for synergistically controlling Tetranychus urticae with a pyrethroid medicament, wherein the Neoseiidae is pyrethroid-resistant;
the resistance of the neoseiulus barkeri to the bromocyanochrysanthester medicament is incomplete dominant polygenic inheritance and is obtained by cultivating the neoseiulus barkeri by the method of the embodiment 1; inoculating tetranychus urticae collected in the field on indoor planted cowpea seedlings, propagating for 2 weeks under appropriate conditions, and counting the number of adult mites and nymph mites on each seedling;
the experimental protocol was set up as follows,
in this example, the pyrethroid drugs were tested in 8 groups, and the experimental protocol was as follows:
the treatment 1 is that sensitive strains of the neoseiulus barkeri bred indoors are respectively connected to cowpea seedlings with tetranychus urticae according to the benefit-to-pest ratio of 1: 10;
treating 2, respectively inoculating the resistance strains of the indoor bred neoseiulus barkeri to the cowpea seedlings with the tetranychus urticae according to the benefit-to-pest ratio of 1: 10;
the treatment 4 is that sensitive strain neoseiulus barkeri is firstly put in according to the proportion of the treatment 1, and then the 3000 times of the selected 2.5 percent deltamethrin emulsifiable solution is applied for foliar spraying;
treatment 5, firstly throwing in the drug-resistant strain neoseiulus barkeri according to the proportion of the treatment 2, and then applying 3000 times of the selected 2.5% deltamethrin emulsifiable solution to carry out foliar spraying;
treatment 6 is to independently select 3000 times of cyhalothrin emulsifiable concentrate foliar spray group of 2.5 percent;
the treatment 7 is that sensitive strain neoseiulus barkeri is firstly put in according to the proportion of the treatment 1, and 3000 times of the selected 2.5 percent cyhalothrin missible oil is applied to carry out foliar spraying after the putting;
the treatment 8 is that the neoseiulus barkeri of the drug-resistant strain is firstly put in according to the proportion of the treatment 2, and 3000 times of the selected cyhalothrin emulsifiable solution with 2.5 percent is applied to carry out foliar spraying after the putting;
the contrast is plants which are grown with tetranychus urticae and are not controlled at all, each plant is a repetition, and 10 repetitions are set for each treatment;
the experimental method comprises the following steps: putting each plant in a plastic tray, isolating the tray with water, processing by a scheme of 1-8, performing base number investigation, performing investigation every 4d after processing, respectively recording the number of tetranychus urticae koch and small neoseiulus barkeri on each cowpea seedling, and performing investigation for 22 d; the test is carried out in a closed insect breeding room, the temperature is 25 +/-1 ℃, the relative humidity is 60-80%, and the illumination L: D is 16: 8.
The experimental results of example 3 are shown in table 1:
TABLE 1 Combined control effect of pyrethroid anti-mite, pyrethroid sensitive strain of neoseiulus barkeri and pyrethroid insecticide on Tetranychus urticae
The above base numbers are the average number of mites per strain, and the different lower case letters indicate that the statistical analysis of each column was significantly different at the 5% level.
Table 1 shows the effect of controlling Tetranychus urticae Koch alone and in combination with 2.5% deltamethrin emulsifiable concentrate and 2.5% cyhalothrin emulsifiable concentrate.
The results show that the control effects of the single use of deltamethrin, cyhalothrin, neoseiulus pasteurianus anti (sensitive) strain and the combined action of deltamethrin (cyhalothrin) and neoseiulus pasteurianus anti (sensitive) strain on tetranychus urticae are greatly different:
after the treatment, the control effect of the 2d, 2.5 percent deltamethrin missible oil, 2.5 percent cyhalothrin missible oil and the two medicaments used together with the small neoseiulus barkeri is obvious (P <0.05) and is higher than the control effect of the single release of the small neoseiulus barkeri resistance (sensitive) strain;
10d, the control effect of the single released neoseiulus barkeri anti (sensitive) strain on tetranychus urticae is not obviously different from that of other treatments;
at the 22d, the control effects of the treatment groups for independently releasing the small neoseiulus barkeri resistant strain and jointly controlling the tetranychus urticae by the two medicaments and the small neoseiulus barkeri resistant strain are both 100 percent, and the control effect is remarkably higher (P <0.05) than that of the control effects of the single application of the deltamethrin and the cyhalothrin (the control effects are respectively 96.20 percent and 96.63 percent) and the combined action of the two medicaments and the small neoseiulus barkeri sensitive strain (the control effects are respectively 95.98 percent and 96.93 percent);
after the two medicaments of 2.5 percent deltamethrin emulsifiable and 2.5 percent cyhalothrin emulsifiable are independently applied, the control effect on the tetranychus urticae on the 2 nd day is 93.18 percent and 94.16 percent respectively, the combined control effect of the two insecticides and the sensitive strain neoseiulus barkeri is 93.73 percent and 95.02 percent respectively, and the combined control effect of the two insecticides and the drug-resistant neoseiulus barkeri strain respectively reaches 97.59 percent and 97.94 percent; the data analysis result shows that the combined control effect (P <0.05) of the pesticide and the neoseiulus barkeri resistant strain is more remarkable than that of the pesticide and the pesticide used alone and the neoseiulus barkeri sensitive strain; the control effect of the two pesticides applied independently and the two pesticides respectively combined with the neoseiulus barkeri sensitive strain has no significant difference (P ≧ 0.05);
the control effect of the agent for treating tetranychus urticae singly from 6d to 10d is not significantly different from the control effect of the agent and the combined application of the neoseiulus barkeri and the neoseiulus barkeri (P ≧ 0.05).
Results of 14d control data show: although the control effect of the agent for independently treating the tetranychus urticae koch and the control effect of the agent and the combined application of the agent and the small new amblyseius barkeri have no significant difference (P is not less than 0.05), the survival number of the tetranychus urticae koch is 0 by the combined control of the two agents and the drug-resistant strain of the small new amblyseius barkeri, and the control is relatively thorough as no increase occurs until the 22 nd day; the two insecticides are applied independently and are respectively combined with the neoseiulus barkeri sensitive strain for application, although the control effect is better from the 2 nd to the 10 th after the control, the control is not thorough, and the generation amount of the tetranychus urticae koch from the 14 th to the 22 th shows a gradually rising trend; therefore, the two medicaments are respectively combined with the neoseiulus barkeri drug-resistant strain to prevent and treat tetranychus urticae koch to achieve better prevention and treatment effects. The control cycle (14 days) of the combined use of the deltamethrin or cyhalothrin new small amblyseius drug-resistant strain is shortened by 57 percent compared with the control cycle (22 days) of the single use of deltamethrin or cyhalothrin, the reduction of the control cycle can greatly reduce the loss of economic crops, and the economic effect is obvious.
In order to prove that the method for synergistically preventing and treating tetranychus urticae by using small neoseiulus barkeri and pyrethroid medicaments is feasible, a large number of experiments are carried out in the subject group, and part of experimental processes and data are shown as follows:
The results of the breeding of the drug resistance of deltamethrin to neoseiulus barkeri are shown in tables 2 and 3. Selecting sublethal measurement of 0.14mg/L as initial concentration of small Sublyseius barkeri resistance breeding according to virulence determination result of original sensitive population (SS) in laboratory, controlling mortality of small Sublyseius barkeri at 79.01-61.22% in 24 hours under breeding of deltamethrin medicament with the concentration, screening each generation later, controlling mortality of small Sublyseius barkeri at 52.11-86.32% by using medicament concentration, breeding for 30 generations, and finally increasing the medicament concentration to 30mg/L, wherein deltamethrin is used for LC of small Sublyseius barkeri50The value is improved from 0.32mg/L to 72.44mg/L, and the drug resistance multiple is 226.38 times. According to the fact that the LD-P lines of RR and SS shown in FIG. 1 are basically parallel and the intercept values are close, it is proved that two populations of small neoseiulus pasteurianus are pure.
TABLE 2 Breeding of neoseiulus barkeri on deltamethrin resistant strains
Note: fs is a sensitive strain of neoseiulus barkeri, and Fn is a resistant strain of neoseiulus barkeri continuously bred.
TABLE 3 virulence determination of different populations of neoseiulus barkeri
Note that a is the dominance value
A large number of experimental studies have found that: the method is characterized in that the method comprises the steps of selecting a drug-resistant strain of the neoseiulus barkeri deltamethrin, carrying out continuous multi-generation high-frequency selection on the concentration of a sub-lethal medicament acting on a population, wherein the mortality rate of the population cannot exceed 85% after each selection, and the too high mortality rate can cause the population not to be easily and quickly recovered, so that the drug-resistant level of the population is reduced in the long recovery process.
The genetic analysis method adopted by the invention is as follows:
1.1 predatory mite bioassay method
The bioassay method is mainly based on the classical conventional standardization method of the spider mites, the glass-dipping method, established by FAO (FAO, 1980). 5-6 concentration gradients are set for testing pesticides in resistance breeding virulence determination, resistance genetic analysis is carried out, the series concentration is increased to 10, and clear water is used as a control. The double-sided adhesive tape is cut into 2 cm and pasted on a glass slide, female adult mites with the same size are selected by a fine hair pen, the back of the female adult mites is pasted on the double-sided adhesive, the female adult mites lie on the back, the feet and mouth can move, each glass slide is pasted with 3 rows, 10 heads of each row, and no mechanical damage is guaranteed. The insect-bearing slides were then immersed in different concentration gradient solutions for 5 seconds, each treating 30 mites, 3 replicates. After the slide is taken out, the redundant liquid drops are sucked by filter paper, the slide is placed on a slide rack and is placed under the appropriate temperature and humidity condition, and after 24 hours, the death condition is checked under the mirror by using a brush pen or a needle.
1.2 Cross-backcross test
Setting orthogonal and inverse cross tests: in order to ensure that the female adult mites subjected to the hybridization test are not mated, 300 heads of female anterior nymphs of the resistant and sensitive populations of small Sublyseius barkeri are picked and raised separately, and after the female adult mites develop, the adult mites are mated with male mites of the resistant and sensitive populations, namely (RR female and multiplied by SS) and (SS female and multiplied by RR), so as to obtain the first filial generation [ F1(F1RS,F1SR)](ii) a Backcross test, pick F1Generation F1RSAnd F1SRBefore femaleIf the mites are independently raised, the mites are backcrossed with parents respectively to obtain F after the mites develop into female mites2Generation, i.e., [ BC ]1(RS♀×RR♂)]And [ BC ]1′(SR♀×SS♂)]. To parents and F respectively1、F2The female adult mites of the generations were bioassayed (see 1.2 for determination) and statistical analysis was performed using the log dose-mortality probability line (LD-P).
1.3 measurement of dominance degree
The apparent and recessive common dominance degree (D) of pest drug resistance inheritance is used for distinguishing, and a D value calculation formula is as follows:
wherein X1、X2、X3LC representing parental homozygote R, heterozygote RS or SR and perceptual homozygote S respectively50A logarithmic value. The resistance gene is completely recessive when D is-1; resistance gene is not fully recessive when-1 < D < 0; 0 < D < 1 is incomplete dominance of the resistance gene; when D is 1, the resistance gene is completely dominant.
1.4 maternal genetic analysis method
Computational analysis was performed using the following dominance analysis of variance formula.
If two Var(D) If the difference is not significant, the resistance is considered to be chromosomal genetic.
1.5 determination of Single and double Gene inheritance
According to the single gene genetic expectation hypothesis proposed by mendelian, calculation of the corresponding frequency of backcrossed individuals under different dose treatments was done according to the following formula (Georghiou and Garber, 1965):
BC1(F1RS♀×RR♂),XV＝W(F1RS)×0.5+W(RR)×0.5 (3)
BC1′(F1SR♀×SS♂),XV＝W(F1SR)×0.5+W(SS)×0.5 (4)
XVdenotes that at a certain concentration, F2Expected mortality of generations, W being F1RS、F1SRMortality of the RR and SS populations at the corresponding concentrations, these values can all be obtained from the LD-P lines of the different populations. From the above values, F can be obtained2Surrogate dose-expected response curve (i.e. at different agent doses, F)2Generation expected mortality maps).
According to the Ka-Square test formula of Sokal et al (1995):
chi-square detection herein (2Test) is used to detect F2The difference between actual dose-response and dose-expected response. F is the actual mortality at a dose, p is the expected mortality at this dose, q is 1-p, n is the total number of tested worms (90) at this dose, r is the number of groups of doses (r 10 in this trial), if F2Substitute of reality2Value and expectation2There is a difference between the values, indicating that the actual situation is different from the expected hypothesis, and is multigenic inheritance, and conversely, is monogenic inheritance.
The genetic analysis of drug resistance results are shown below:
(1) dominance degree analysis result
Calculating to obtain F according to the dominance degree measurement formulaISRDegree of dominance DSRValue 0.8938, FISRDegree of dominance DRSA value of 0.7617, both of which are 0<D<1, indicating that the resistance gene of the neoseiulus barkeri to the deltamethrin is incompletely dominant inheritance.
(2) Genetic analysis results of maternal lines
The result of the calculation according to the formula of the dominance variance shows that Var (D)RS) The value is 0.0024, Var (D)SR) A value of 0.0015, based on
Is calculated to
The 95% confidence limits for the D values obtained using D + -2 SE calculations are shown in Table 3, and overlap between the two results indicates that there is no significant difference between the RS and SR lines from the F1 cross, indicating that the resistance genes are on the autosome and that the maternally effects are less influential.
(3) Determination of monogenic or polygenic inheritance
FIGS. 2 and 3 are F2Surrogate dose-actual, expected response LC-P line, as can be seen from the results in the figure, regardless of F2(FISRMale parent is X RR male parent or F2(FISRAnd (3) obviously separating an actual curve and an expected curve of the male parent and the female parent, indicating that the actual reaction LC-P line and the expected LC-P line are different, and preliminarily judging that the resistance inheritance of the neoseiulus barkeri to the deltamethrin is polygenic inheritance. Chi-square test is shown in Table 4, the actual observed chi-square value (X) is calculated2) Is obviously greater than the expected chi-square value, and further proves that the resistance of the neoseiulus barkeri to the deltamethrin is controlled by multiple genes.
TABLE 4 backcross F2Chi-square test for substitute observed value and expected value
| Backcross F2Substitute for Chinese traditional medicine | Desired chi-square value | Actual chi-square value | Degree of freedom |
| F2(FISR♀×RR♂) | 15.51 | 49.08 | 8 |
| F2(FISR♀×SS♂) | 15.51 | 136.39 | 8 |
Table 5 is the cross-resistance spectrum of neoseiulus pasteurii deltamethrin resistant and sensitive strains to several different agents. The results show that the neoseiulus pasteurianus resistant strain produced different degrees of resistance to the 13 pyrethroid insecticides tested. The resistance level to cyhalothrin is highest, and the resistance multiple is 2210.91 times. The resistance multiple to deltamethrin is 226.38. The cross-resistance to other tested insecticides was bifenthrin, chlorpyrifos, lambda-cyhalothrin, ethofenprox, beta-cypermethrin, etoxazole in sequence from high to low with multiples of resistance 146.40, 110.71, 91.82, 80.64, 30.56, 20.43, and 17.58, respectively. No cross-resistance was developed to carbosulfan and pyridaben.
TABLE 5 Cross-resistance spectra of Amblyseius barkeri deltamethrin resistant and sensitive strains to different agents
Experiment 4 determination of developmental history between resistant and sensitive strains of neoseiulus barkeri
Placing sponge (diameter 4.5cm and thickness 0.5cm) and absorbent cotton (diameter 4.0cm) in a culture dish with diameter 5cm from bottom to top, placing in a stainless steel tray (50cm × 35cm), and adding distilled water to soak the sponge and absorbent cotton to form a floating shape;
cutting cowpea leaves damaged by Tetranychus urticae, making into dish with diameter of 2.5cm (the density of Tetranychus urticae is maintained at 40-60 heads/dish, and supplement can be performed when the quantity is insufficient), and placing the back of the leaf on absorbent cotton;
respectively picking eggs laid by the Amblyseius barkeri for less than 6h, placing single eggs into a prepared leaf disc, and feeding the eggs in a single way under the ambient condition of room temperature (25 ℃, RH 80-90% and photoperiod 14L: 10D); the treated samples were 100 specimens. Record the status of each mite and death day by day (1 record each 8:00 am and 20:00 pm); after the female adult mites emerge for 1d, 1 male adult mite in the same age is selected and paired, and the initial sample size is not less than 60. The oviposition and survival number of female adult mites are observed and counted every day, prey (Tetranychus urticae) is supplemented in time, distilled water is added into a culture dish regularly, and the leaf discs are replaced within an interval period of 6-8 d.
The development cycle of the neoseiulus barkeri resistant and sensitive strain is shown in Table 6. The egg stage, the young mite stage, the later nymph stage and the days for completing one generation of the neoseiulus pasteurii deltamethrin resistant strain have no significant difference with the sensitive strain, and the former nymph stage and the pre-spawning stage of the neoseiulus pasteurii deltamethrin resistant strain are significantly longer than the sensitive strain (P < 0.05).
TABLE 6 Bromopsis Bromophila, neoseiulus pasteurii, Bromophrin resistant and sensitive strains in development cycle
| Stage of development | Sensitive strain | Resistant strains |
| In ovo (d) | 1.55±0.14 | 1.67±0.11 |
| Mite stage (d) | 0.78±0.05 | 0.83±0.03 |
| Front if mite period (d) | 1.76±0.10* | 1.34±0.14 |
| Postnymous period (d) | 1.56±0.12 | 1.53±0.18 |
| In the early stage of spawning (d) | 3.38±0.14* | 3.08±0.14 |
| Generation (d) | 12.61±0.39 | 13.10±0.48 |
The data in the table are mean ± sem; data from the same row showed significant differences at 0.05 level (t test)
As can be seen from Table 7, the egg laying period, the daily egg production and the total egg production of the neoseiulus barkeri deltamethrin resistant population are all higher than those of the sensitive population, wherein the variation of the day of the egg laying period and the total egg production of each female is significant (P < 0.05). The egg hatchability of the deltamethrin resistant neoseiulus pasteurianus is slightly lower than that of the sensitive population, but the difference is not obvious.
TABLE 7 Productivity of neoseiulus barkeri deltamethrin resistant and sensitive strains
| Sensitive strain | Resistant strains | |
| Spawning period (Tian) | 20.34±0.57 | 23.62±1.36* |
| Egg laying amount per female per day (granule/female) | 1.54±0.06 | 1.60±0.03 |
| Every female always lay eggs (granules) | 31.09±3.11 | 35.22±2.06* |
| Hatching rate (%) | 95.47±1.88 | 94.66±3.78 |
The data in the table are mean ± sem; data from the same row indicates a significant difference at 0.05 level (t-test).
Claims (10)
1. A method for preventing and controlling two-spotted spider mite by the synergy of Amblyseius barkeri and pyrethroid medicaments is characterized in that: the neoseiulus barkeri is the neoseiulus barkeri with pyrethroid resistance.
2. The method for synergistically controlling tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments as claimed in claim 1, wherein the method comprises the following steps: the drug resistance of the neoseiulus barkeri to the deltamethrin medicament is incomplete dominant polygenic inheritance.
3. The method for synergistically controlling tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments as claimed in claim 1, wherein the method comprises the following steps: the pyrethroid medicaments are cyhalothrin, deltamethrin, bifenthrin, lambda-cyhalothrin, ethofenprox, beta-cypermethrin and cypermethrin; etoxazole as a miticide; the organophosphorus chemical chlorpyrifos; any one of the second generation new nicotinoyl insecticides thiamethoxam.
4. The method for synergistically controlling tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments as claimed in claim 1, wherein the method comprises the following steps: and (3) inoculating the small neoseiulus barkeri with pyrethroid resistance to the vegetable seedlings with the tetranychus urticae, spraying the pyrethroid pesticide on the leaf surfaces of the vegetable seedlings, and performing field management.
5. The method for synergistically controlling tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments, as claimed in claim 4, is characterized in that: inoculating small neoseiulus barkeri with pyrethroid resistance to the vegetable seedling with Tetranychus urticae at a ratio of 1:5-20, and spraying onto the leaf surface of the vegetable seedling at a concentration of 5 × 10-3-10×10-3Performing field management on g/L cyhalothrin or deltamethrin.
6. The method for synergistically controlling tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments, as claimed in claim 5, is characterized in that: inoculating small neoseiulus barkeri with pyrethroid resistance to the vegetable seedling with tetranychus urticae at a ratio of 1:10, and spraying 2.5% of effective components on the leaf surface of the vegetable seedlingPerforming field management on Fujing emulsifiable solution or 3000 times of a 2.5% deltamethrin emulsifiable solution diluent; the application concentration of cyhalothrin or deltamethrin is 8.3 × 10-3g/L。
7. The method for synergistically controlling tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments as claimed in claim 1, wherein the method comprises the following steps: the breeding method of the small neoseiulus barkeri with pyrethroid resistance comprises the following steps:
picking out female adult mite of Amblyseius barkeri from parent, and applying 98% deltamethrin crude drug to LC of female adult mite25The mites are treated by dosage, and after 24 hours, the surviving neoseiulus barkeri is transplanted into the pink mites; meanwhile, in the process of feeding the flour mites, deltamethrin agents with the same concentration are sprayed in the feed, the agent spraying concentration is gradually increased along with the increase of drug resistance of flour mite populations, the new small amblyseius barkeri is fed by the drug-resistant flour mites, and the steps are repeated and screened for multiple generations.
8. The method for the synergistic control of Tetranychus urticae by Amblyseius barkeri and pyrethroid medicaments according to claim 7, which is characterized in that: the breeding method of the small neoseiulus barkeri with pyrethroid resistance comprises the following steps:
selecting LC25The dose of 0.14mg/L is used as the initial concentration of the resistance breeding of the small neoseiulus barkeri, under the breeding of the deltamethrin medicament with the concentration, the mortality of the small neoseiulus barkeri in 24 hours is between 79.01 and 61.22 percent, each generation of screening is carried out, the concentration of the medicament is used to control the mortality of the small neoseiulus barkeri to be between 52.11 and 86.32 percent, and the small neoseiulus barkeri with pyrethroid resistance is obtained by breeding for 30 generations.
9. The method for synergistically controlling tetranychus urticae by using neoseiulus barkeri and pyrethroid medicaments as claimed in claim 1, wherein the method comprises the following steps: the breeding method of the small neoseiulus barkeri with pyrethroid resistance comprises the following steps:
taking a plurality of culture dishes with the diameter of 5cm, sequentially placing sponge and absorbent cotton from bottom to top, placing the culture dishes on a stainless steel tray, and adding distilled water to soak the sponge and the absorbent cotton to form a slightly floating state;
cutting cowpea leaves damaged by Tetranychus urticae, making into a leaf disc with diameter of 2.5cm, keeping the density of Tetranychus urticae at 40-60 heads/disc, supplementing when the number is insufficient, and placing the back of the leaf upwards on absorbent cotton;
respectively picking eggs laid by the Amblyseius barkeri for less than 6h, placing single eggs into the prepared leaf disc, and feeding the eggs in a single way under the environmental conditions of room temperature 25 ℃, RH 80-90% and photoperiod 14L: 10D.
10. A small neoseiulus barkeri with pyrethroid resistance is characterized in that: obtained by breeding according to the method of claim 7 or 8.
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