CN110881464A

CN110881464A - Attractant formula for preventing and treating Holotrichia parallela imagoes in North China

Info

Publication number: CN110881464A
Application number: CN201911237288.3A
Authority: CN
Inventors: 尹姣; 李克斌; 李晓峰; 张帅; 梅向东; 伊洛森
Original assignee: Institute of Plant Protection of Chinese Academy of Agricultural Sciences
Current assignee: Institute of Plant Protection of Chinese Academy of Agricultural Sciences
Priority date: 2019-12-05
Filing date: 2019-12-05
Publication date: 2020-03-17

Abstract

The invention discloses an attractant formula for controlling Holotrichia parallela imago in North China in the technical field of biological control of pests. The attractant formula consists of cis-3-hexenyl acetate and cis-9-octadecenyl acetate, and the proportion of each component is 100: 11.3. The attractant formula for controlling the adult Holotrichia parallela has good control effect on the Holotrichia parallela, and the Holotrichia parallela cannot generate resistance; the preparation process is simple, the preparation cost is low, the preparation method is harmless to human and livestock, and the preparation method is environment-friendly and has important significance for comprehensively treating Holotrichia parallela in North China.

Description

Attractant formula for preventing and treating Holotrichia parallela imagoes in North China

Technical Field

The invention belongs to the technical field of biological control of pests, and particularly relates to an attractant formula for controlling Holotrichia parallela imago in North China.

Background

Holotrichia oblita of Holotrichia of the order Coleoptera belongs to the family Chinemorphae of the order Coleoptera and is an important underground pest in dry farming areas of North China. Its larvae (grubs) eat the underground parts of the plant, often causing severe yield loss in peanuts, soybeans, potatoes, etc. As grubs have soil inhabitation and are hidden in harmful mode, the grubs are mainly controlled by chemical pesticides with long lasting period in the past, such as hexachloro-cyclohexane soprocide, aldicarb and the like, and the pesticides have high toxicity and high residue and are banned at home for a long time. And with the abuse of chemical pesticides, such pesticides have caused a serious social problem. A new measure is to achieve the aim of reducing grubs from controlling the quantity of adults and reducing the egg laying amount of the adults, namely a soil insect prevention and control strategy for 'soil insect control on the ground'. The developed attractant has the advantages of high efficiency and environmental friendliness in controlling the adult population of Holotrichia parallela in production, and the ingredients of the formula of the attractant mainly include plant volatile matters, insect aggregation pheromones, sex pheromones and the like.

Disclosure of Invention

The invention aims to provide an attractant formula for controlling Holotrichia parallela imagoes in North China, so as to improve the effect of comprehensively controlling Holotrichia parallela in North China.

The method selects plant volatile matters, Holotrichia parallela gathering pheromone and sex pheromone, obtains the optimal attracting concentration of each compound monomer through a chemotactic behavior reaction test, mixes the three compound monomers pairwise according to the optimal attracting concentration, and respectively adopts 17 formulas of the plant volatile matters + the gathering pheromone, the plant volatile matters + the sex pheromone and the gathering pheromone + the sex pheromone. The active formula is gradually screened through an antenna potential reaction and a chemotaxis behavior reaction test, the formula is compared with a single component for attracting effect, and the attracting effect of the formula is tested through an indoor cage attracting test.

The 5 plant volatile components are dibutyl phthalate, cinnamaldehyde, cis-3-hexenyl acetate, 1-hexanol and cis-3-hexenyl isobutyrate; 2 kinds of aggregation pheromone components are cis-9-octadecenal and cis-9-octadecenyl acetate; the 1 sex pheromone component is glycine methyl ester. The prepared 17 groups of attractant formulas are screened step by step, 3 groups of attractant formulas are found to have better effects, and the results are better when the 3 groups of attractant formulas are subjected to cage trapping effect tests, wherein one group of attractant formulas has the best effect. The composition and content of these 3 formulations are shown in table 1.

TABLE 1

The optimal formula comprises cis-3-hexenyl acetate and cis-9-octadecenyl acetate.

Preferably, the ratio of the 2 components of cis-3-hexenyl acetate and cis-9-octadecenyl acetate is 100: 11.1.

1. According to the preparation method of the attractant formula for controlling the adult Holotrichia parallela, the optimal attracting concentration of 3 pheromone monomers, namely cis-9-octadecenal, cis-9-octadecenyl acetate and sex pheromone methyl glycine, of the Holotrichia parallela aggregated pheromone to the adult Holotrichia parallela is determined through a chemotactic behavior reaction test (the optimal attracting concentration of another 5 plant volatile matter monomers is known), based on the optimal attracting concentration, 100 mu L of cis-3-hexenyl acetate and 11.1 mu L of cis-9-octadecenyl acetate are determined, and the attractant components are mixed and dissolved in 100mL of paraffin oil solvent.

The application of the attractant formula for controlling Holotrichia parallela imago is used for controlling Holotrichia parallela.

Compared with the prior art, the invention has the following excellent effects: the attractant formula for controlling the adult Holotrichia parallela has good control effect on the Holotrichia parallela, and the Holotrichia parallela cannot generate resistance; the preparation method is simple, the preparation cost is low, the method is harmless to human and livestock, the environment is friendly, and the method has important significance for comprehensively treating the Holotrichia parallela in North China.

Drawings

Fig. 1 and 2 show the insect antenna potential test graphs of the 17-group attractant formula for the male and female Holotrichia parallela in North China, respectively.

FIG. 3 shows the trend behavior of Holotrichia parallela against 6 formulas. The number of the vertical axis is the formula number. Asterisks indicate that the tropism of the Holotrichia parallela imago to different formulas is remarkably different from that of a control (paraffin oil) (P is less than 0.05), and double asterisks indicate that the extremely remarkable difference is achieved (P is less than 0.01) (chi square test).

Figure 4 shows a graph comparing the attractant effect of 3 formulations with their individual components. The numbering of the left longitudinal axis is the formula number, and the right side is the single component. Asterisks indicate that the tropism of the Holotrichia parallela imago to different formulas is remarkably different from that of a single component of the Holotrichia parallela imago (P is less than 0.05), and double asterisks indicate that the extremely remarkable difference is achieved (P is less than 0.01) (chi square test).

FIG. 5 shows the cage trapping effect of 3 formulas on Holotrichia parallela adults in North China. The number of the longitudinal axis is the formula number. Asterisks indicate that the tropism of the Holotrichia parallela imago to different formulas is remarkably different from that of a control (paraffin oil) (P is less than 0.05), and double asterisks indicate that the extremely remarkable difference is achieved (P is less than 0.01) (chi square test).

Detailed Description

The present invention will be described in detail below with reference to the drawings and examples, but the practice of the invention is not limited thereto.

Example 1 screening of pheromone monomer for optimal attraction concentration of Holotrichia parallela in North China

The optimal attracting concentration of 5 plant volatile monomer on the Holotrichia parallela imago is known, wherein the concentrations of dibutyl phthalate, cinnamaldehyde, cis-3-hexenyl acetate, 1-hexanol and cis-3-hexenyl isobutyrate are respectively 0.11 mu g/mu L, 0.33 mu g/mu L, 1.00 mu g/mu L, 0.33 mu g/mu L and 3.00 mu g/mu L.

The attraction concentrations of the Holotrichia parallela pheromone monomers in North China are respectively determined. 3 compounds of gathering pheromone cis-9-octadecenal, acetic acid cis-9-octadecenyl ester and sex pheromone glycine methyl ester are respectively diluted into 5 concentration gradients of 0.11 mu g/ml, 0.33 mu g/ml, 1 mu g/ml, 3 mu g/ml, 9 mu g/ml and the like by taking paraffin oil as a solvent, and the optimal attraction concentration is searched through a chemotactic behavior reaction test. The test is scheduled to be carried out at 20:30-23:30 according to the diurnal and nocturnal habits of the test insects. The measuring room is clean and clean, the ventilation condition is good, the temperature is 25 +/-1 ℃, and the humidity is kept at about 70%. By using paraffin oil as a reference, 20 μ L of compound monomers with different concentrations as treatment were uniformly spread on 4cm × 3mm (length × width) filter paper strips, and then two side arms of a Y-shaped tube were placed as an odor source. 1 test insect subjected to starvation treatment for 24 hours is connected to a position 3cm away from the main arm, and if the test insect crawls to more than 1/2 of any side arm and the retention time is more than 1min, the test insect is considered to have reaction; and if the test insects still stay on the main arm or turn around to leave the Y-shaped pipe after 10min, the test insects are considered to be non-reactive. Each treatment tested 30 females and males, which were repeated 3 times. When the Y-shaped pipe is replaced and treated every time, 75% alcohol and distilled water are respectively used for washing the Y-shaped pipe, and the Y-shaped pipe can be continuously used after being dried. The calculation formula of the selective activity in the Y-type olfactometer test is as follows:

reaction rate (%) - (the number of insects tested on the source arm + the number of insects tested on the control arm) ÷ the number of insects tested × 100%;

the reaction rate (%) was selected as the number of insects tested on the source arm ÷ (number of insects tested on the source arm + number of insects tested on the control arm) × 100%.

The test results are shown in table 2.

TABLE 2

Data in table are mean ± sem; different letters after the same column of data indicate significant differences at a P <0.05 level as tested by Duncan's new double-pole-difference method.

As can be seen from Table 2, the reaction rate of all treatments was higher than 87.78%, indicating that the test insects were active and the results were reliable. The aggregation pheromone cis-9-octadecenal, the acetic acid cis-9-octadecenyl ester and the sex pheromone glycine methyl ester of the Holotrichia parallela have the strongest effect of attracting test insects when the concentrations are 0.11 mu g/mu L, 0.11 mu g/mu L and 3 mu g/mu L, and the selective reaction rates respectively reach 60.12 percent, 66.09 percent and 64.00 percent.

Example 2 formulation of attractants

The selected plant volatile matter, the aggregation pheromone and the sex pheromone of the Holotrichia diomphalia Bates are mixed pairwise to obtain 17 groups of binary attractant formulas. The content proportion of each component in the formula is determined according to the result of the optimal attracting concentration of each compound monomer to the test insects, so that the concentrations of the two compound monomers in the formula are the optimal attracting concentrations. 100mL of each formula was prepared using paraffin oil as the solvent. The components and contents of each formulation are shown in table 3.

TABLE 3

Example 3 determination of the antennal potential of test insects for different formulations

Active test insects are selected from the insect breeding box, the base part of the antenna is lightly pressed by tweezers to open 3 cheek pieces of the antenna, the antenna is quickly cut off from the base part and is fixed at two ends of the electrodes by conductive adhesive, and the antenna between the two electrodes cannot be stained with the adhesive, otherwise, the test result can be influenced. And (3) uniformly coating 20 mu L of the formula solution on a filter paper strip with the thickness of 4cm multiplied by 3mm by using a liquid transfer gun, longitudinally folding the filter paper strip, putting the folded filter paper strip into a Pasteur tube, sealing, and taking the filter paper strip for use in the test. During the test, the laboratory is ensured to be clean and free from peculiar smell, and no factors influencing the test result, such as personnel walking, are caused. And debugging software, and performing odor stimulation on the antennae after the base line is stable. The continuous flow rate was 500 mL/min and the stimulation flow rate was 40 mL/min. Each stimulation time is 0.2s, and the interval between two stimulations is more than 30 s. Test formulations were randomly drawn and tested, all with 20 μ L paraffin oil as a control. Cis-3-hexenol was used as standard reference volatiles. Female and male adults were tested separately. Each formulation and standard reference volatiles were tested 3 times in duplicate, and each replicate was replaced with a new filter paper strip to avoid the effect of concentration deviation on the assay results, and a paraffin oil control was tested once for each formulation and standard reference volatiles. Repeat 8 antennae. Each antenna tested 10 formulations. The calculation formula of the relative value of the reaction of measuring the antenna potential is as follows:

the relative antennal potential response (%) - (treatment antennal potential response-control antennal potential response) ÷ (reference antennal potential response-control antennal potential response) × 100%

The test results are shown in fig. 1 and fig. 2.

It can be seen that the relative response values of the Holotrichia parallela female in North China to 6 species of the

formulas

2B, 6F, 7G, 9I, 10J and 11K are all over 5722% and are all significantly different from those of the other formulas (P is less than 0.05). In this formulation, the highest relative values of reaction were found for formulation 2B up to 11093%, and for

formulations

6F, 7G, 9I, 10J, 11K up to 7537%, 6759%, 5981%, 5796%, 5722%, respectively.

The relative reaction values of the andropathia chinensis benth male worms to 7 types of the

formulas

2B, 6F, 7G, 9I, 10J, 11K and 14N are larger, are all more than 3566 percent and are all obviously different from the relative reaction values of other formulas (P is less than 0.05). Of these 7 formulations, the highest relative value of reaction was formulation 10J, up to 4712%. The

formulas

2B, 14N, 9I, 6F, 11K and 7G respectively reach 4677%, 4438%, 3983%, 3878%, 3809% and 3566%.

From the relative response values, the female and male responses were more consistent to the 17 formulations, but the female had a greater relative response value than the male. The 6

formulas

2B, 6F, 7G, 9I, 10J and 11K can cause stronger electrophysiological reactions of males and females, so that the 6 formulas can be used for further screening.

Example 4 test insects' chemotactic behavior response to different formulations

The test was carried out as in example 1 for the determination of the chemotactic behavior of 6 formulations, each of which tested 30 male and female Holotrichia parallela adults from North China, and repeated 3 times. Further screening the formulation for behavioral reactivity against the test insect.

The test results are shown in fig. 3 and table 4.

TABLE 4

It can be seen that the attracting amount of the

formulations

9I, 10J and 11K to the test insects is much more than that of the control (P <0.01), and the attracting test insects of the 3 formulations and the control are respectively 118 (formulation 9I) and 49 (CK), 119 (formulation 10J) and 49 (CK), 121 (formulation 11K) and 49 (CK). The luring amount of the

formulas

2B, 6F and 7G to the test insects is not different from that of the control (P is more than 0.05). The reaction rate of the test insects to 6 formulas is higher than 91.11%, the test insect activity is stronger, the test result is reliable, and the selective reaction rate is between 52.35% and 71.21%. Wherein, the highest selective reaction rate is formula 11K which reaches 71.21 percent, and then formula 10J and 9I, and the selective reaction rates respectively reach 70.84 percent and 70.72 percent. The selective reaction rates of the three formulas are small in difference and have no significant difference, but the selective reaction rates are all significantly higher than those of other formulas (P < 0.05). Therefore, the formulations screened by the taxis behavior response measurement are 3 formulations in total, namely 9I, 10J and 11K.

Example 5 comparison of attraction Effect of different formulations with their Individual Components

The procedure is as in example 1, determined by a chemotactic behavior response test. 9I, 10J and 11K are compared with two compound monomer components respectively for attracting effects. One arm of the Y-tube was filled with 20. mu.L of the formulation solution, and the other arm was filled with 20. mu.L of the single component (at a concentration consistent with the monomer concentration of the compound in the formulation), and each treatment was repeated 3 times for 30 male and female Holotrichia parallela adults.

The results are shown in FIG. 4.

As can be seen, the number of the test insects induced by the formula 9I and the cis-3-hexenyl acetate is 95 and 66 respectively, and the difference between the two is significant (P < 0.05); the number of the test insects induced by the formula 9I and the cis-9-octadecenyl acetate is 96 and 71 respectively, and the difference between the two is not significant (P is more than 0.05). The number of the test insects induced by the formula 10J and the 1-hexanol is 98 and 62 respectively, and the difference between the two is very obvious (P is less than 0.01); the number of test insects induced by the formula 10J and the cis-9-octadecenyl acetate is 103 and 61 respectively, and the two are also very different (P is less than 0.01). The number of the test insects induced by the formula 11K and the dibutyl phthalate is 94 and 69 respectively, and the difference between the two is remarkable (P is less than 0.05); the number of the test insects attracted by the formula 11K and the glycine methyl ester is 103 and 65 respectively, and the difference between the two is very obvious (P is less than 0.01).

Thus, formula 9I has a remarkable synergistic effect on cis-3-hexenyl acetate, but has no remarkable synergistic effect on cis-9-octadecenyl acetate; the formula 10J shows extremely remarkable synergistic effect on 1-hexanol and cis-9-octadecenyl acetate which are single components; formula 11K has a significant synergistic effect on dibutyl phthalate, and has a very significant synergistic effect on glycine methyl ester. The 3 formulations were subjected to the next cage trapping effect test.

EXAMPLE 63 cage trap Effect of formulations on test insects

Formulations

9I, 10J, 11K were tested for cage trapping effect in 1 m.times.1 m glass cages. A foam plate is placed in the glass cage at a position 15cm away from the bottom, and the area of the foam plate is the same as that of the bottom of the glass cage. 4 flowerpots (the upper diameter is 18cm, the lower diameter is 13cm and the height is 13cm) are respectively placed at two ends of two diagonal lines of the bottom surface of the glass cage, and the upper edges of the flowerpots are parallel and level to the foam board. The thickness of soil in the flowerpot is 10cm, the small lure core is inserted into a 1000 mu L liquid-transfering gun head to form a combined body, and then the combined body is inserted into the soil in the flowerpot, so that the small lure core is 3cm higher than the soil. The experiment arrangement is carried out at 20:00-23:00, two flowerpots on one diagonal line of a glass cage are used as treatment groups, and 60 mu L of the same formula solution is added into small lure cores; the other two diagonal flowerpots are used as a control group, and the small lure of the control group is added with 60 mu L of paraffin oil. 30 male and female test insects are put into each glass cage respectively. The laboratory is kept clean and clean, the temperature is 25 +/-1 ℃, and the humidity is kept at about 70%. The trapping results were observed after 1 h. If the test insects fall into the flowerpot or climb on the small lure core, the reaction is considered; if the test insects do not enter the flowerpot, no reaction is considered. After the counting was completed, the glass cage was opened for ventilation, and the test was repeated after 0.5h, with 3 repetitions of each formulation. The cage trapping test has the calculation formula as follows:

the reaction rate (%) (number of tested insects in the flowerpot of the treatment group + number of tested insects in the flowerpot of the control group)/number of tested insects is multiplied by 100%

The reaction rate (%) was selected as the number of the test insects in the treated flowerpot/(the number of the test insects in the treated flowerpot + the number of the test insects in the control flowerpot) × 100%

The test results are shown in fig. 5 and table 5.

TABLE 5

As can be seen, the insect attracting number of the 3 formulas and the insect attracting number of the control are respectively 126 heads and 45 heads (9I), 116 heads and 60 heads (10J), 115 heads and 64 heads (11K). The insect-attracting number of the insect-attracting population is very different from that of a control (P < 0.01). The reaction rate of the Holotrichia parallela imago to the three formulas is higher than 95%, which indicates that the test insects are more active and the test result is reliable. The highest reaction rate was chosen for formulation 9I, reaching 74.06%. The

formulations

10J and 11K were followed, and the selectivity to the reaction rates reached 65.89% and 64.25%, respectively. The cage trap test result shows that the formula 9I has a strong attraction effect on the Holotrichia parallela imagoes in North China.

Claims

1. An attractant formula for controlling Holotrichia parallela imago in North China, which is characterized in that the attractant formula comprises cis-3-hexenyl acetate and cis-9-octadecenyl acetate.

2. The attractant formulation for controlling adult Holotrichia parallela according to claim 1, wherein the ratio of the 2 components of cis-3-hexenyl acetate and cis-9-octadecenyl acetate is 100: 11.3.

3. The preparation method of the attractant formula for controlling adult Holotrichia parallela according to claim 1, wherein the optimal attracting concentrations of 3 pheromone monomers, namely cis-9-octadecenal, cis-9-octadecenyl acetate and sex pheromone glycine methyl ester, to the adult Holotrichia parallela are determined through a chemotactic response test, and based on the optimal attracting concentrations of the other 5 plant volatile matter monomers, 100 μ L of cis-3-hexenylacetate and 11.1 μ L of cis-9-octadecenyl acetate are determined, and the attractant components are mixed and dissolved in 100mL of paraffin oil solvent.

4. The use of the attractant formulation for controlling adult Holotrichia parallela according to claim 1, wherein the attractant formulation is used for controlling Holotrichia parallela.