Solenopsis invicta biological control agent with repelling effect, behavior inhibition effect and poisoning activity and application

Abstract

The invention belongs to the field of biological control, and particularly relates to a solenopsis invicta biological control agent with an avoidance effect, a behavior inhibition effect and poisoning activity and application. Fumigation activity tests and the like find that the exocarpium citri grandis essential oil has good fumigation activity on solenopsis invicta, the sand digging amount, attack frequency and climbing force of the solenopsis invicta can be reduced, and the exocarpium citri grandis essential oil has good avoidance effect, behavior inhibition effect and poisoning activity, can be used as a solenopsis invicta biological prevention and control agent to be applied to prevention and control of the solenopsis invicta, and has good application prospects. Good application prospects are realized.

Description

Technical Field

[0001] The invention belongs to the field of biological control, and specifically relates to a biological control agent for red imported fire ants with repellent effect, behavior inhibition effect and toxic activity and application thereof. Background Art

[0002] The red fire ant (Solenopsis invicta Buren) is a dangerous invasive organism with serious harm. It eats crops in large quantities, plunders food resources, and has almost no natural enemies. It can build nests and holes in fields, hilly woodlands, urban communities, highway gardens, etc., which seriously threatens the safety of humans and animals and the ecological environment. How to protect humans and animals from harm by red fire ants in normal production and life is an urgent problem to be solved in the current prevention and control of red fire ants. At present, the use of chemically synthesized pesticides to poison red fire ants is the main means of prevention and control. Due to the single means of prevention and control and limited prevention and control effects, it cannot play a good protective role for humans and animals. It is urgent to develop new environmentally friendly and safe red fire ant control agents for humans and animals.

[0003] Essential oils are extracted from plant-specific aromatic substances. They are easily degraded in the natural environment, are not likely to cause environmental pollution, contain rich active ingredients, are safe for humans and animals, and often exhibit novel modes of action such as repelling and avoiding pests, providing possibilities for the development of new pest control agents and innovative pest control methods. However, there are few reports on the use of essential oils to control red fire ants, and even fewer studies on the use of such modes of action as avoidance to control red fire ants.

[0004] Therefore, an essential oil having a repellent effect on red fire ants is found and processed into a product, which can be used in daily production and life to repel red fire ants, thereby protecting humans and animals from harm by red fire ants. Summary of the invention

[0005] In order to overcome the deficiencies and shortcomings of the prior art, the primary purpose of the present invention is to provide an application of tangerine peel essential oil in the prevention and control of red fire ants.

[0006] Another object of the present invention is to provide the use of the above-mentioned tangerine peel essential oil in repelling red fire ants.

[0007] Another object of the present invention is to provide the use of the above-mentioned tangerine peel essential oil in inhibiting the behavior of red fire ants.

[0008] The fourth object of the present invention is to provide the use of the above-mentioned tangerine peel essential oil in poisoning red fire ants.

[0009] A fifth object of the present invention is to provide a biological control agent for red imported fire ants having repellent effect, behavior inhibition effect and toxic activity.

[0010] The purpose of the present invention is achieved through the following technical solutions:

[0011] Application of tangerine peel essential oil in the prevention and control of red fire ants;

[0012] The Huazhou pomelo essential oil is an essential oil extracted from Huazhou pomelo tissues and organs by at least one extraction method such as distillation, freeze-pressing extraction, oil separation, solvent extraction, immersion, and supercritical carbon dioxide extraction;

[0013] The tissue or organ in question is at least one of a flower, a leaf, a fruit, a branch, a peel, etc.;

[0014] The Huazhou pomelo peel essential oil is preferably an essential oil extracted by steam distillation from Huazhou pomelo peel as a raw material;

[0015] The tangerine peel essential oil is preferably prepared by the following method:

[0016] The peel of young Huazhou pomelo fruit is mixed with water, and then the essential oil is extracted by steam distillation;

[0017] The material-liquid ratio of the steam distillation method is preferably 1:10;

[0018] The extraction conditions of the steam distillation method are preferably a distillation temperature of 105°C and a distillation time of 3h;

[0019] The tangerine peel essential oil preferably comprises the following components: limonene, trans-oxidized linalool, γ-terpinene, α-terpineol, 4-terpene alcohol, myrcene, β-cubane and cis-oxidized linalool, and the contents of the above components decrease in sequence;

[0020] The tangerine peel essential oil preferably contains the following components in relative amounts: 25-26% limonene, 11-12% trans-oxidized linalool, 7-8% γ-terpinene, 7-8% α-terpineol, 6-7% 4-terpene alcohol, 6-7% myrcene, 5-6% β-cubane, and 5-6% cis-oxidized linalool;

[0021] The tangerine peel essential oil further preferably contains the following components in relative amounts: 25.80% limonene, 11.04% trans-oxidized linalool, 7.94% γ-terpinene, 7.89% α-terpineol, 6.35% 4-terpene alcohol, 6.15% myrcene, 5.88% β-cubane, and 5.29% cis-oxidized linalool;

[0022] The application of the tangerine peel essential oil in repelling red fire ants.

[0023] The use of the tangerine peel essential oil in inhibiting the behavior of red fire ants;

[0024] The behavior includes at least one of sand digging, attacking, and climbing;

[0025] The application of the tangerine peel essential oil in poisoning red fire ants;

[0026] A biological control agent for red imported fire ants with repellent effect, behavioral inhibition effect and toxic activity, comprising the above-mentioned tangerine peel essential oil;

[0027] A method for preventing and controlling red fire ants comprises the following steps:

[0028] Spraying the above-mentioned tangerine peel essential oil or the preparation containing the above-mentioned tangerine peel essential oil directly on the area where red fire ants need to be controlled;

[0029] The areas where red fire ants need to be controlled include farmland, plantations, ant nests, human bodies, etc.

[0030] Compared with the prior art, the present invention has the following advantages and effects:

[0031] (1) The present invention found through fumigation activity test that the essential oil of tangerine peel has good fumigation activity against red imported fire ants. 50 The value was 0.0213 μL / mL, which was significantly higher than other plant volatile components (sweet orange essential oil, citronella essential oil, rose essential oil, limonene). Further GC-MS component analysis found that although the main components of tangerine peel essential oil and sweet orange essential oil all contain limonene, the LC value of tangerine peel essential oil was 0.0213 μL / mL. 50 The value was significantly lower than that of other plant volatile components (sweet orange essential oil, citronella essential oil, rose essential oil), especially lower than that of pure limonene. The result shows that the composition and ratio of the main components in essential oils directly affect the toxic effect on red fire ants. The other main components of tangerine peel essential oil and limonene may have a synergistic promoting effect, thereby improving its fumigation activity.

[0032] (2) The present invention further studies the effect of tangerine peel essential oil on the sand digging behavior, attack behavior and climbing behavior of red fire ants. The test results show that compared with other volatile components (sweet orange essential oil, citronella essential oil, DEET, rose essential oil, limonene), tangerine peel essential oil can reduce the amount of sand digging, the number of attacks and the climbing ability of red fire ants, that is, it has a good avoidance effect and behavior inhibition effect, indicating that it has a potential protective effect on the protected object.

[0033] (3) The present invention discovers for the first time that tangerine peel essential oil has good repellent effect, behavior inhibitory effect and toxic activity against red fire ants, and can be used as a biological control agent for red fire ants in the prevention and control of red fire ants.

[0034] (4) Citrus aurantium essential oil has a pleasant fragrance, is safe and harmless to humans and animals, is easily degraded in the natural environment, and is not easy to pollute the environment. Its application in the prevention and control of red fire ants, especially the protection of field workers, to avoid harm from red fire ants provides a novel and environmentally friendly solution and method with good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 This is the ion flow chart of GC-MS analysis of tangerine peel essential oil.

[0036] Figure 2 This is a graph showing the effects of different tested volatile components on the sand-digging behavior of red fire ants.

[0037] Figure 3 It is a bubble chart showing the effects of different tested volatile components on the attack intensity of red fire ants.

[0038] Figure 4 This is a graph showing the effects of different volatile components on the dynamic climbing ability of red fire ants.

[0039] Figure 5 This is a trend chart showing the effects of different tested volatile components on the rate of reduction in the climbing ability of red fire ants. DETAILED DESCRIPTION

[0040] The present invention is further described in detail below in conjunction with embodiments and drawings, but the embodiments of the present invention are not limited thereto.

[0041] The experimental reagents and test insects involved in the embodiments are as follows:

[0042] (1) Orange essential oil, citronella essential oil, and rose essential oil were provided by Guangzhou Fuling Xinji Biotechnology Co., Ltd., and limonene (purity 95%) and DEET (purity 99%) were purchased from Shanghai MacLean Biochemical Technology Co., Ltd.

[0043] (2) The red fire ants tested in this embodiment were collected from a citrus demonstration base near the north gate of the Fruit Research Institute of Guangdong Academy of Agricultural Sciences. The collected ant nests were placed in a plastic bucket (10 L), and a "water test tube" was placed in the bucket to provide water for the ant colony. Mealworms were used as food, and then the nests were stably built for 2 days under the conditions of a light cycle of 12L:12D, a temperature of 26±3°C, and a relative humidity of 70±5%.

[0044] Example 1

[0045] (1) Extraction of tangerine peel essential oil

[0046] Fresh Huazhou pomelo young fruits (5 cm in diameter) were harvested, the peel (including the white sponge layer) was peeled, 200 g of the peel was put into a flask, water was added at a solid-liquid ratio of 1:10, and then the flask was placed in a distillation apparatus to extract essential oil by steam distillation, wherein the extraction conditions were: distillation temperature 105°C, boiler temperature 110°C, and distillation time 3h; the extracted Huazhou orange red essential oil was stored in the dark, and the essential oil extraction rate was calculated according to the following formula:

[0047] Essential oil extraction rate (%) = (essential oil mass / raw material mass) × 100

[0048] (2) Identification of components of tangerine peel essential oil

[0049] The tangerine peel essential oil obtained in step (1) was subjected to GC-MS analysis, and the specific analysis conditions were: injection volume of 1 μL, carrier gas of high-purity helium (99.999%), split ratio of 50:1, constant flow mode, flow rate of 1 mL / min. The temperatures of the injection port, ion source and transmission line were 250°C, 260°C and 280°C, respectively. The GC heating program was as follows: 40°C for 3 min; then heated to 160°C at a rate of 3°C / min and retained for 1 min; then heated to 200°C at a rate of 5°C / min and retained for 1 min; finally heated to 240°C at a rate of 8°C / min and retained for 3 min. The MS conditions were as follows: EI (electron impact) ion source, electron impact energy of 70 eV, positive ion scanning mode, mass scanning range m / z 45-400 amu.

[0050] After testing, in this embodiment, the extraction rate of tangerine peel essential oil is 1.36%. Figure 1 This is the electron flow chart of GC-MS analysis of tangerine peel essential oil. Figure 1 Qualitative and relative quantitative analysis were performed, and the results are shown in Table 1. As can be seen from Table 1, the main components of the tangerine peel essential oil prepared in this embodiment are limonene, trans-oxidized linalool, γ-terpinene, α-terpineol, 4-terpene alcohol, myrcene, β-cubane, and cis-oxidized linalool, and their contents are 25.80%, 11.04%, 7.94%, 7.89%, 6.35%, 6.15%, 5.88%, and 5.29%, respectively.

[0051] Table 1 GC-MS analysis of the main components of tangerine peel essential oil

[0052]

[0053]

[0054] Example 2 Fumigation activity of tangerine peel essential oil against red imported fire ants

[0055] In this example, the toxicity of essential oils to red imported fire ants was evaluated by fumigation activity, and the specific method is as follows:

[0056] (1) Take a 150 mL conical flask (actual volume is 227 mL), place a 1.5 mL centrifuge tube filled with pure water at the bottom of the flask to provide water for the worker ants (sealed with cotton wool), and place 1.5 cm long mealworms as food; evenly apply a circle of talcum powder on the inner wall near the flask mouth;

[0057] (2) Then, 20 red imported fire ant workers (body length about 3-4 mm) that had been starved for 3 h were introduced into the conical flask; PCR tubes with a volume of 0.1 mL were taken, and different volumes (weights were recorded at the same time) of the volatile components to be tested (citrus essential oil, sweet orange essential oil, citronella essential oil, DEET, rose essential oil or limonene prepared in Example 1) were added respectively, and one end of the PCR tube was tied tightly with a thin thread, and the other end was tied to the center of a toothpick slightly longer than the outer diameter of the conical flask mouth, so that the PCR tube containing the volatile components to be tested was suspended in the center of a 150 mL conical flask (actual volume was 227 mL), and the bottom of the PCR tube was about 4 cm away from the bottom of the conical flask, and then the conical flask mouth was sealed with plastic wrap, and 3 replicates were performed for each treatment;

[0058] (3) After 24 hours of fumigation, the number of live ants in each triangular bottle was counted, the mortality rate was calculated, and the lethal concentration (LC) of the corresponding volatile components to red fire ants was calculated by the least squares method. 50 ).

[0059] The results are shown in Table 2. The LC 50 The value is 0.0213 μL / mL (μL: essential oil volume, mL: container volume), the LC of sweet orange essential oil 50 The value is 0.0624μL / mL, citronella essential oil is 0.0485μL / mL, rose essential oil is 0.465μL / mL, and limonene is 0.056μL / mL. From the results, it can be concluded that the fumigation activity of tangerine peel essential oil against red fire ants is second only to DEET, which is about 2.930 times that of sweet orange essential oil, 2.27 times that of citronella essential oil, 21.831 times that of rose essential oil, and 2.630 times that of limonene, indicating that tangerine peel essential oil has a strong toxic activity against red fire ants; in addition, the main component of tangerine peel essential oil is limonene, but its fumigation activity against red fire ants is significantly higher than that of pure limonene, indicating that other main components of tangerine peel essential oil and limonene may have a synergistic promoting effect, thereby improving its fumigation activity; similarly, the main component of sweet orange essential oil is also limonene, but its fumigation activity is lower than that of pure limonene and tangerine peel essential oil, which to a certain extent indicates that there is no synergistic promoting effect between its main component limonene and other main components, thereby making its fumigation activity higher than that of pure limonene.

[0060] Table 2 Fumigation activity of different tested volatile components against red imported fire ants

[0061]

[0062] Example 3 Effect of Citrus aurantium essential oil on the behavior of red fire ants

[0063] (1) Effects of tangerine peel essential oil on the sand-digging behavior of red imported fire ants

[0064] ① Take a 9cm plastic culture dish, and use a soldering iron to drill a hole about 3cm from the center of the bottom to make it just big enough to put in a 2mL centrifuge tube. While it is still heated, use force to immediately push the 2mL centrifuge tube through the culture dish until the tube is flush with the bottom of the culture dish. Fix another centrifuge tube in the same way on the other side of the dish. Weigh 20g of sterilized fine sand (passed through a 30-mesh sieve), moisten it with 1.9mL of sterilized pure water, add 100μL of acetone solution (containing 20μL of the volatile components to be tested), mix well and put it into a 2mL centrifuge tube; the control (CK) group uses 100μL of acetone; in addition, fix two centrifuge tubes filled with water in the vertical direction of the center of the straight line where the centrifuge tubes of the treatment group and the control group are located to provide water for the red fire ant workers;

[0065] ② Then, 23 to 26 red fire ant workers (body length of about 4 to 5 mm) that had been starved for 3 hours were inoculated into the center of the culture dish. After 24 hours, the red fire ants in the control group and the essential oil treatment group were collected to dig out the sand, and the deaths of the red fire ants in the centrifuge tubes and culture dishes were checked and recorded, and the mortality rate was calculated (mortality rate = number of dead red fire ant workers / total number of inoculated red fire ant workers × 100%); 5 replicates were set for each treatment.

[0066] Results Figure 2 and Table 3, among which, the weight of sand dug out by red fire ants after treatment with the 6 tested volatile components is from small to large: DEET < tangerine peel essential oil < citronella essential oil < limonene < sweet orange essential oil < rose essential oil. Except for rose essential oil, the other tested volatile components can cause the death of worker ants, and the mortality rate is: DEET > tangerine peel essential oil > citronella essential oil > limonene > sweet orange essential oil. For tangerine peel essential oil, the weight of sand dug out by the control group is 4 times that of the essential oil group, indicating that tangerine peel essential oil significantly affects the behavior of red fire ants digging sand and shows a strong avoidance effect on red fire ants. At the same time, the mortality rate of red fire ant workers is 34.89%, further indicating that tangerine peel essential oil has good toxicity activity against red fire ants.

[0067] Table 3 Effects of different tested volatile components on the sand excavation of red imported fire ant workers

[0068]

[0069] (2) Effects of tangerine peel essential oil on the attack behavior of red fire ants

[0070] ①Use a 9cm diameter petri dish and place two mealworms symmetrically with the center point 2cm away from the edge. Apply 10μL of the volatile components to be tested evenly on the surface of one of them, and leave the other one untreated.

[0071] ② Place 20 red fire ant workers (about 4-5 mm in length) that have been starved for 3 hours in the middle of the culture dish, then cover the dish and seal it with plastic wrap. Place it under a high-definition camera and count the number of attacks by red fire ant workers on mealworms every hour for a total of 4 hours, with 5 replicates for each treatment.

[0072] The results are shown in Table 4. In the 0-1h period, it was found that the red fire ant workers attacked the yellow mealworms in the tangerine peel treatment group, and the number of attacks at this time was 25% of the control group. In the 1-2h period, it was observed that the number of attacks by the red fire ant workers in the tangerine peel essential oil treatment group was 14.29% of the control group. In the 2-3h period, it was observed that the number of attacks by the red fire ant workers in the tangerine peel essential oil treatment group was 8% of the control group. In the subsequent records, no red fire ant workers were observed to attack the yellow mealworms in the treatment group. Tangerine peel essential oil and DEET have a significant inhibitory effect on the aggressive behavior of red fire ant workers, and the inhibitory effect is significantly enhanced with the increase of treatment time. The ability of these five ingredients to affect the aggressive behavior of worker ants is in the following order: DEET>tangerine peel essential oil>citronella essential oil>sweet orange essential oil>limonene. Among them, rose essential oil has almost no effect on the aggressive behavior of worker ants.

[0073] In order to further evaluate the attack behavior and ability of the red fire ant workers, the present invention further analyzes the changes in the dynamic attack intensity of the red fire ant workers, wherein the attack intensity = the number of attacks in the treatment group / the percentage of the number of attacks in the control group × 100%, Figure 3 It is a bubble chart of the changes in the attack intensity of different volatile components tested in different time periods. It can be seen from the figure that the attack intensity of red fire ant workers gradually decreased to zero after the treatment of tangerine peel essential oil and DEET; after the treatment of sweet orange essential oil and citronella essential oil, the attack intensity first decreased and then increased with the increase of time, while the attack intensity of red fire ant workers slightly decreased within 1 to 4 hours after the treatment of limonene.

[0074] Table 4 Effects of different tested volatile components on the attack times of red imported fire ants

[0075]

[0076] (3) Effect of tangerine peel essential oil on the climbing ability of red imported fire ants

[0077] ① Red imported fire ant worker ants (body length of about 4 to 5 mm) that had been starved for 3 hours were used as the research objects, and the fumigation activity test method of Example 2 was used as the reference, wherein the ants were divided into 4 groups according to the fumigation treatment time (fumigation 1 hour group, fumigation 2 hours group, fumigation 3 hours group, fumigation 4 hours group), and each group had 2 treatments (control group and test volatile component treatment group, each treatment was set with 5 replicates).

[0078] ② When the scheduled fumigation time is up, remove the plastic wrap, insert a small bamboo stick and observe the climbing ability of each worker ant. If the height of the climbing bamboo stick is not less than 2cm, it is considered to have climbing ability.

[0079] The results of the effects of different volatile components on the climbing ability of red fire ant workers are shown in Table 5. As can be seen from Table 5, all red fire ants in the control group have good vitality and good climbing ability. However, the behavioral ability of red fire ant workers in the treatment groups of tangerine peel essential oil, sweet orange essential oil, citronella essential oil, DEET and limonene was significantly affected, and weakened with the extension of treatment time. The effects of these five volatile components on the climbing ability of red fire ant workers are as follows: DEET> tangerine peel essential oil> limonene> citronella essential oil> sweet orange essential oil> rose essential oil.

[0080] In order to further evaluate the climbing behavior and ability of red fire ant workers, the present invention further analyzed the changes in the dynamic climbing ability of red fire ant workers, wherein the climbing ability of the control group is 1, and the climbing ability of the treatment group = the number of red fire ant workers with climbing ability in the treatment group / the number of red fire ant workers with climbing ability in the control group × 100%. Figure 4 . For tangerine peel essential oil, the climbing ability of red fire ant workers began to weaken significantly at 1 hour, which was 13.00% weaker than the control group; at 2, 3, and 4 hours, it was 23%, 42%, and 64% weaker than the control, respectively, indicating that tangerine peel essential oil can significantly and permanently reduce the behavioral ability of red fire ant workers, thereby reducing the harm to people. Although sweet orange essential oil, citronella essential oil, and limonene can also reduce the climbing ability of red fire ant workers, the effect is significantly weaker than tangerine peel essential oil and DEET.

[0081] In order to further evaluate the changing trend of the climbing ability of the red fire ant workers, the present invention further analyzed the reduction speed / trend of the climbing ability of the red fire ant workers (climbing ability (N h)-climbing ability (N+1h)). The results are shown in Figure 5 .from Figure 5 It can be seen that after treatment with tangerine peel essential oil and DEET, the climbing ability of red fire ants decreased faster, followed by limonene, and rose essential oil had the least effect on the climbing ability of red fire ants. It is worth noting that between 2 and 3 hours, the climbing ability of red fire ants decreased faster after treatment with tangerine peel essential oil than with DEET.

[0082] Table 5 Effects of different tested volatile components on the climbing ability of red imported fire ant workers

[0083]

[0084] Note: The above data are the number of worker ants with climbing ability (head).

[0085] The above embodiments are preferred implementation modes of the present invention, but the implementation modes of the present invention are not limited to the above embodiments. Any other changes, modifications, substitutions, combinations, and simplifications that do not deviate from the spirit and principles of the present invention should be equivalent replacement methods and are included in the protection scope of the present invention.

Claims

1. An application of tangerine peel essential oil in the prevention and control of red fire ants.

2. The use according to claim 1, characterized in that: The Huazhou pomelo essential oil is obtained by taking Huazhou pomelo tissues and organs as raw materials and extracting the essential oil through at least one of distillation, freeze-pressing extraction, oil separation, solvent extraction, immersion and supercritical carbon dioxide extraction.

3. The use according to claim 1, characterized in that: The Huazhou pomelo peel essential oil is an essential oil extracted from Huazhou pomelo peel by steam distillation.

4. The use according to claim 3, characterized in that: The tangerine peel essential oil is prepared by the following method: The peel of young Huazhou pomelo fruit is mixed with water, and then the essential oil is extracted by steam distillation; The solid-liquid ratio of the steam distillation method is 1:10; The extraction conditions of the steam distillation method are as follows: a distillation temperature of 105° C. and a distillation time of 3 hours.

5. The use according to claim 1, characterized in that: The tangerine peel essential oil comprises the following components: limonene, trans-oxidized linalool, gamma-terpinene, alpha-terpineol, 4-terpene alcohol, myrcene, beta-cubane and cis-oxidized linalool, and the contents of the above components decrease in sequence.

6. An application of tangerine peel essential oil in repelling red fire ants.

7. An application of tangerine peel essential oil in inhibiting the behavior of red fire ants.

8. An application of tangerine peel essential oil in poisoning red imported fire ants.

9. A biological control agent for red imported fire ants having repellent effect, behavior inhibition effect and toxic activity, characterized in that Contains Citrus aurantium essential oil.

10. A method for preventing and controlling red fire ants, comprising the following steps: Spray the tangerine peel essential oil or a preparation containing the tangerine peel essential oil directly on the area where red fire ants need to be controlled.

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