FENGYCIN FAMILY LIPOPEPTIDES IN PEST CONTROL

Abstract

An application of fengycin family lipopeptides in pest control is provided, and Fengycin family lipopeptides are used for pest control, with especially strong killing effects on Homoptera and Coleoptera pests. A method is provided for treating seeds with fengycin to protect seeds and plant organs formed later from pests, so as to protect seeds from pests by coating the fengycin family lipopeptides on the surface of seeds.

Claims

1. An application of fengycin family lipopeptides in pest control.

2. The application according to claim 1, wherein the fengycin family lipopeptides are one of fengycin A, fengycin B, fengycin C, fengycin S, plipastatin A and plipastatin B, or more than one of fengycin A, fengycin B, fengycin C, fengycin S, plipastatin A and plipastatin B as a mixture.

3. The application according to claim 1, wherein the pests are Homoptera pests or Coleoptera pests.

4. The application according to claim 3, wherein the Homoptera pests are one or more of Aphis craccivora Koch, Aphis gossypii, Brevicoryne brassicae and Macrosiphum avenae.

5. The application according to claim 3, wherein the Coleoptera pests are one or more of Protaetia brevitarsis, Holotrichia parallela, Anomala corpulenta and Polyphylla gracilicornis.

6. The application according to claim 1, wherein the application is to prepare the fengycin family lipopeptides into liquid medicine and then treat plants or soil to control pests.

7. A method for protecting seeds and later-formed plant organs from pests, comprising treating seeds with the fengycin family lipopeptides according to claim 1.

8. The method according to claim 7, wherein a mass concentration of the fengycin family lipopeptides is 0.1-100 μg/milliliter (ml).

9. The method according to claim 7, wherein the seeds are coated with a coating agent containing the fengycin family lipopeptides.

10. The method according to claim 9, wherein the coating agent further comprises a film forming agent or an adhesive.

11. An improved method of controlling pests, the method comprising application of a fengycin family lipopeptides.

12. The improvement of claim 11, wherein the fengycin family lipopeptides are selected from a group consisting of: fengycin A; fengycin B; fengycin C; fengycin S; plipastatin A; and plipastatin B.

13. The improvement of claim 11, wherein the fengycin family lipopeptides are selected from or more than one of fengycin A, fengycin B, fengycin C, fengycin S, plipastatin A and plipastatin B as a mixture.

14. The improvement of claim 11, wherein the pests are Homoptera pests or Coleoptera pests.

15. The improvement of claim 14, wherein the Homoptera pests are one or more of Aphis craccivora Koch, Aphis gossypii, Brevicoryne brassicae and Macrosiphum avenae.

16. The improvement of claim 14, wherein the Coleoptera pests are one or more of Protaetia brevitarsis, Holotrichia parallela, Anomala corpulenta and Polyphylla gracilicornis.

17. The improvement of claim 11, wherein the fengycin family lipopeptides comprises a liquid medicine treated on seeds, plants or soil.

18. The improvement of claim 17, wherein the liquid medicine has a mass concentration of the fengycin family lipopeptides of 0.1-100 μg/milliliter (ml).

Description

DETAILED DESCRIPTION OF THE EMBODIMENTS

[0021] Now various exemplary embodiments of the present application will be described in detail. This detailed description should not be taken as a limitation of the present application, but should be understood as a more detailed description of some aspects, characteristics and embodiments of the present application.

[0022] It should be understood that the terms mentioned in the present application are only used to describe specific embodiments, and are not used to limit the present application. In addition, for the numerical range in the present application, it should be understood that each intermediate value between the upper limit and the lower limit of the range is also specifically disclosed. Every smaller range between any stated value or the intermediate value within the stated range and any other stated value or the intermediate value within the stated range is also included in the present application. The upper and lower limits of these smaller ranges can be independently included or excluded from the range.

[0023] Unless otherwise stated, all technical and scientific terms used herein have the same meanings commonly understood by those of ordinary skill in the field to which this application relates. Although the present application only describes preferred methods and materials, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present application. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and/or materials related to the documents. In case of conflict with any incorporated documents, the contents of this specification shall prevail.

[0024] Without departing from the scope or spirit of the present application, it is obvious to those skilled in the art that many modifications and changes may be made to the specific embodiments of the present specification. Other embodiments obtained from the description of the present application will be obvious to the skilled person. The description and embodiment of that application are only exemplary.

[0025] Unless otherwise specified, the experimental methods used in the embodiments are conventional experimental methods in the field; Materials and reagents used in the embodiments can be obtained from commercial sources unless otherwise specified.

Embodiment 1

[0026] Preparation of Fengycins

[0027] Bacillus amyloliquefaciens WS-8 (preservation number: CGMCC No. 11787) is used for fermentation to prepare fengycins. The specific steps are as follows:

[0028] S1, carrying out amplification culture on the bacterial strain to obtain fermentation liquor;

[0029] S2, centrifuging the fermentation liquor to remove thallus to obtain sterile supernatant;

[0030] S3, adding amberlity XAD-7HP macroporous resin to the sterile supernatant for adsorption, and shaking at 4° C. for 12 hours;

[0031] S4, filtrating and collecting the resin, washing it with distilled water for three times, then with 60% ethanol (volume/volume), and concentrating the eluent to obtain a crude extract;

[0032] S5, using high performance liquid chromatography for further purification, including, loading mobile phase A, which is acetonitrile containing 0.1% (volume ratio) trifluoroacetic acid, and mobile phase B, which is ultrapure water containing 0.1% (volume ratio) trifluoroacetic acid, into WondaSil C18(4.6 mm×150 mm) chromatographic column with a particle size of 5 μm, using 10%-90% acetonitrile for linear gradient elution with the flow rate 1 mL/min, detecting the wavelength as 214 nm and the column temperature as 30° C., collecting the components corresponding to 30-50 min, and using the mass spectrometry (Table 1) to confirm that fengycins are obtained.

TABLE-US-00001 TABLE 1 Mass spectrometry detection results of purified products Compound serial m/z m/z Characteristic Determination number [M + H]+ [M + Na]+ fragmention result 1 1435.7660 1457.7474 540.77 C.sub.14 Fengycin A 2 1449.7848 1471.7635 540.77 C.sub.15 Fengycin A 3 1463.7975 1485.7784 554.79 C.sub.14 Fengycin B 4 1449.7836 1471.7632 540.77 C.sub.15 Fengycin A 5 1449.7816 1471.7626 540.77 C.sub.15 Fengycin A 6 1477.8138 1499.7944 554.79 C.sub.15 Fengycin B 7 1463.7969 1485.7777 540.77 C.sub.16 Fengycin A 8 1463.7974 1485.7789 540.77 C.sub.16 Fengycin A 9 1477.8138 1499.7888 540.77 C.sub.17 Fengycin A 10 1477.8140 1499.7932 540.77 C.sub.17 Fengycin A 11 1477.8157 1499.7963 540.77 C.sub.17 Fengycin A 12 1491.8285 1513.8091 554.79 C.sub.16 Fengycin B 13 1505.8439 1527.8250 554.79 C.sub.17 Fengycin B 14 1447.8036 1469.7852 540.77 C.sub.15:1 Fengycin A 15 1461.8197 1483.8000 540.77 C.sub.16:1 Fengycin A 16 1475.8328 1497.8135 554.79 C.sub.15:1 Fengycin B

Embodiment 2

[0033] Insecticidal Effect of the Fengycins on HOMOPTERA Insects

[0034] Dissolving the fengycins in methanol to prepare 200 μg/mL mother liquor, and then diluting the mother liquor with water to 5 μg/mL, 10 μg/mL and 20 μg/mL liquid medicine, and using clean water treatment as a control; soaking young broad bean stems in insecticidal experiment of Aphis craccivora Koch, young cotton stems in insecticidal experiment of Aphis gossypii, young cabbage leaves in insecticidal experiment of Brevicoryne brassicae, young stems of wheat in insecticidal experiment of Macrosiphum avenae, soaking the young broad bean stems, the young cotton stems, the young cabbage leaves and the young stems of wheat in the above-mentioned liquid medicine for 10 minutes, then absorbing water with filter paper, taking them out, putting them in a plate with filter paper, adding 2 mL of deionized water to keep moisture, then covering the plate with fresh-keeping film, puncturing holes, and adding 1 mL of water every 12 hours; testing 50 aphids in each concentration respectively, repeating for 3 times, using a brush head to pick 50 aphids of each species with similar age and size, putting them on the plate to starve for 4 hours, then adding the young broad bean stems, the young cotton stems, the young cabbage leaves and the young wheat stems treated with the above-mentioned liquid medicine onto the plate respectively, changing every 12 hours, culturing overnight in an incubator at 25° C., recording the number of aphids killed, and calculating the mortality rates of aphids at different dilution ratios.

[00001]$\mathrm{The}\mathrm{mortality}\mathrm{rate}=\frac{\mathrm{the}\mathrm{number}\mathrm{of}\mathrm{aphids}\mathrm{killed}}{\mathrm{the}\mathrm{number}\mathrm{of}\mathrm{aphids}\mathrm{for}\mathrm{testing}}⨯100\%$

[0035] Measuring the mortality rates of the aphids: After the aphids are treated overnight, touching the aphids with a brush head, and if the aphid does not move, it is regarded as dead. Count and record the number of dead aphids. It can be seen from the implementation data results in Table 2 that the fengycins have a good control effect on Homoptera insects.

TABLE-US-00002 TABLE 2 Control effect of the fengycins on Homoptera insects Mortality rate (%) Aphis Processing craccivora Aphis Brevicoryne Macrosiphum group Koch gossypii brassicae avenae CK 2.00 1.33 1.33 2.00  5 μg/mL 86.00 87.33 82.00 84.00 10 μg/mL 94.00 94.00 91.33 91.33 20 μg/mL 100.00 98.67 99.33 98.67

Embodiment 3

[0036] Insecticidal Effects of the Fengycins on Coleoptera Insects

[0037] Dissolving the fengycins in methanol to prepare 200 μg/mL mother liquor, and then diluting the mother liquor with water to prepare 5 μg/mL, 10 μg/mL and 20 μg/mL liquid medicine, using the liquid medicine to prepare medicine soil, with the mass ratio of the liquid medicine to soil 1:5, and using medicine soil that prepared with clean water treatment as a control. Using chopped peanuts as food. Using plastic buckets with a capacity of 4 L as containers, and placing 2 L of evenly mixed medicinal soil in each bucket, and adding a proper amount of equal food to mix well. Adopting a medicinal soil method, selecting the grubs of the third instar larvae of Protaetia brevitarsis, Holotrichia parallela, Anomala corpulenta and Polyphylla gracilicornis with the same size, and keeping them in the environment with relative humidity of 50%-70% and temperature of 25° C.-28° C. for 24 hours. Treating 15 third instar larvaes with the different medicine soil prepared above respectively, repeating for 3 times. After inoculating the larvaes, culturing them indoors at normal atmospheric temperature, and counting the mortality rate after 56 hours, as shown in Table 3.

TABLE-US-00003 TABLE 3 Killing effect of the fengycins on different COLEOPTERA larvaes Mortality rate (%) Processing Protaetia Holotrichia Anomala Polyphylla group brevitarsis parallela corpulenta gracilicornis CK 0.00 0.00 0.00 0.00  5 μg/mL 82.22 80.00 82.22 80.00 10 μg/mL 88.89 91.11 88.89 86.67 20 μg/mL 97.78 95.56 93.33 95.56

Embodiment 4

[0038] Dissolving the fengycins in methanol to prepare 200 μg/mL mother liquor, and then diluting the mother liquor with water into 0.1 μg/mL, 0.5 μg/mL, 1 μg/mL, 5 μg/mL, 10 μg/mL, 20 μg/mL, 50 μg/mL and 100 μg/mL liquid medicine to soak peanuts, and using peanuts that soaking in clean water as a control. Using plastic buckets with a capacity of 4 L as containers, and placing 2 L of evenly mixed soil in each bucket, and adding 50 treated peanuts to mix well. Selecting the grubs of the third instar larvae of Holotrichia parallela with the same size, and keeping them in the environment with relative humidity of 50%-70% and temperature of 25° C.-28° C. for 24 hours. Repeating the treatment for 3 times. The number of each test insect in each treatment is 15, repeating the treatment for 3 times. After inoculating the larvaes, culturing them indoors at normal atmospheric temperature, and counting the mortality rate of the larvaes and integrity rate of the peanuts after 56 hours, as shown in Table 4.

TABLE-US-00004 TABLE 4 Insecticidal and protective effects of epiploin on seeds Processing Mortality rate Peanut intactness group (%) rate (%) CK 0.00 8  0.1 μg/mL 53.33 47.33  0.5 μg/mL 64.44 52.00    1 μg/mL 75.56 56.00    5 μg/mL 86.67 62.00   10 μg/mL 95.56 74.00   20 μg/mL 97.78 76.67   50 μg/mL 100.00 79.33  100 μg/mL 100.00 81.33

[0039] The above are only preferred embodiments of the present application, and are not intended to limit the present application. On the premise of not departing from the design spirit of the present application, all kinds of modifications and improvements made by ordinary technicians in the field to the technical schemes of the present application should fall in the scope of protection defined by the claims of the present application.