Use of diethyl fumarate in prevention and treatment of soybean cyst nematode

Abstract

This invention discloses the application of diethyl fumarate in the control of soybean cyst nematodes, belonging to the field of agricultural biotechnology. Through screening the nematicidal activity of fumaric acid and its derivatives, this invention identifies diethyl fumarate as a novel nematicidal compound and provides its application in the control of soybean cyst nematodes. Experimental results show that diethyl fumarate has excellent lethal activity against second-instar larvae of soybean cyst nematodes, with a median lethal concentration (LD50) comparable to that of the commercially available nematicide fluopyram, and can also significantly inhibit the formation of soybean root cysts. Diethyl fumarate has advantages such as low toxicity, easy degradation, and safety, making it suitable for green control and compensating for the shortcomings of existing nematicides. This invention provides a novel, environmentally friendly, and effective pesticide resource for the green control of soybean cyst nematodes.

Description

Technical Field

[0001] This invention relates to the field of agricultural biotechnology, and in particular to the application of diethyl fumarate in the control of soybean cyst nematodes. Background Technology

[0002] Soybean cyst nematode ( Heterodera glycines Soybean cyst nematode (SCN) is one of the most serious soil-borne pathogens affecting soybean production, causing significant yield reductions and even crop failure. Major soybean-producing areas are chronically affected by soybean cyst nematodes, resulting in substantial yield and economic losses and threatening soybean production and revitalization. Currently, production mainly relies on cultivating resistant varieties, crop rotation, and chemical nematicides for control. Commonly used chemical nematicides have problems such as high toxicity, residue risks, and the easy development of resistance. Fumarate esters have been reported for their antibacterial properties due to their simple structure, low toxicity, and good environmental compatibility; however, the nematicidal activity of fumaric acid, diethyl fumarate, and dipropyl fumarate against soybean cyst nematodes has not been publicly reported.

[0003] Currently, conventional nematicides are highly toxic and leave significant environmental residues in the control of soybean cyst nematodes, failing to meet the requirements of green agriculture. Meanwhile, resources of highly effective, low-toxicity, and easily degradable small-molecule nematicides are scarce, limiting the range of options available for production. Furthermore, the activity levels, optimal concentrations, and field application methods of fumaric acid, diethyl fumarate, and dipropyl fumarate against soybean cyst nematodes have not been developed or utilized, leaving a significant technological gap. Summary of the Invention

[0004] The purpose of this invention is to provide the application of diethyl fumarate in the control of soybean cyst nematodes, so as to solve the problems existing in the prior art. Diethyl fumarate has excellent lethal activity against second-instar larvae of soybean cyst nematodes. This invention provides a new type of pesticide resource with clear efficacy and environmental friendliness for the green control of soybean cyst nematodes.

[0005] To achieve the above objectives, the present invention provides the following solution: This invention provides the application of diethyl fumarate in the preparation of nematicides.

[0006] Furthermore, the nematicide is a soybean cyst nematicide.

[0007] The present invention also provides a nematicide with diethyl fumarate as the active ingredient.

[0008] Furthermore, the concentration of the diethyl fumarate is 0.001%-0.004%.

[0009] The present invention also provides the application of diethyl fumarate or the above-mentioned nematicides in the control of soybean cyst nematodes.

[0010] Furthermore, the control of soybean cyst nematode includes killing soybean cyst nematodes and inhibiting the formation of cysts by soybean cyst nematodes.

[0011] Furthermore, the concentration of the diethyl fumarate is 0.001%-0.004%.

[0012] The present invention also provides a method for controlling soybean cyst nematodes, comprising the step of applying diethyl fumarate or the above-mentioned nematicide to the rhizosphere soil.

[0013] Furthermore, the control of soybean cyst nematode includes killing soybean cyst nematodes and inhibiting the formation of cysts by soybean cyst nematodes.

[0014] Furthermore, the concentration of the diethyl fumarate is 0.001%-0.004%.

[0015] The present invention discloses the following technical effects: This invention screens fumaric acid and its derivatives for nematicidal activity, identifying diethyl fumarate as a novel nematicidal compound, and provides its application in controlling soybean cyst nematodes. Experimental results show that diethyl fumarate exhibits excellent lethal activity against second-instar larvae of soybean cyst nematodes, with a median lethal concentration (LD50) comparable to that of the commercially available nematicide fluopyram, and it also significantly inhibits the formation of soybean root cysts. Diethyl fumarate possesses advantages such as low toxicity, easy degradation, and safety, making it suitable for green control and compensating for the shortcomings of existing nematicides. This invention provides a novel, environmentally friendly, and effective pesticide resource for the green control of soybean cyst nematodes. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0017] Figure 1 A statistical chart showing the number of root sporangia in soybeans infected with nematodes after treatment with fumaric acid and diethyl fumarate. Detailed Implementation

[0018] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as a limitation of the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0019] It should be understood that the terminology used in this invention is merely for describing particular embodiments and is not intended to limit the invention. Furthermore, with respect to numerical ranges in this invention, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Any stated value or intermediate value within a stated range, as well as each smaller range between any other stated value or intermediate value within said range, is also included in this invention. The upper and lower limits of these smaller ranges may be independently included or excluded from the range.

[0020] Unless otherwise stated, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. While only preferred methods and materials have been described herein, any methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this invention. All references to this specification are incorporated by way of citation to disclose and describe methods and / or materials associated with those references. In the event of any conflict with any incorporated reference, the content of this specification shall prevail.

[0021] Various modifications and variations can be made to the specific embodiments described in this specification without departing from the scope or spirit of the invention, as will be apparent to those skilled in the art. Other embodiments derived from this specification will also be apparent to those skilled in the art. This specification and embodiments are merely exemplary.

[0022] The terms “include,” “including,” “have,” “contain,” etc., used in this article are all open-ended terms, meaning that they include but are not limited to.

[0023] The experimental materials used in Examples 1 and 2 of this invention are as follows: Test nematode: Soybean cyst nematode ( Heterodera glycines Second instar larva J2; Test compounds: fumaric acid, diethyl fumarate, dipropyl fumarate; Control agent: Fluopyram (commercially available nematicide, positive control); Solvent control: 0.33% ethanol; Blank control: ddH2O.

[0024] Example 1: Determination of the lethal activity of fumaric acid and its derivatives against second-instar larvae J2 of soybean cyst nematode. In this embodiment, fumaric acid and its derivatives were screened for nematicidal activity. A series of concentration solutions were set up, and nematode suspensions were added. After treatment for 24 h and 48 h, the number of dead nematodes was counted and the mortality rate was calculated. The results are shown in Table 1 and Table 2.

[0025] Table 1. 24-h in vitro nematicidal activity of fumaric acid and its derivatives against soybean cyst nematode. Table 1 shows that diethyl fumarate exhibited excellent nematicidal activity within 24 h, with a 100% mortality rate for soybean cyst nematodes at both 0.004% and 0.003% concentrations. At a concentration of 0.002%, the mortality rate reached 88.3±1.9%, and the nematicidal activity of diethyl fumarate was close to that of the positive control fluopyram (94.2±1.4%). It also showed some nematicidal activity at lower concentrations, significantly higher than the blank and solvent controls. Fumaric acid and dipropyl fumarate showed no significant lethal activity against soybean cyst nematode J2 within 24 h.

[0026] Table 2. In vitro nematicidal activity of fumaric acid and its derivatives against soybean cyst nematode after 48 h. Table 2 shows that the nematicidal activity of diethyl fumarate further increased with the treatment time extended to 48 h. At a concentration of 0.002%, the mortality rate of soybean cyst nematodes reached 97.1±1.1%, comparable to the positive control fluopyram (97.7±0.7%). The nematicidal activity was also significantly enhanced at low concentrations, significantly higher than the blank and solvent controls. Dipropyl fumarate only showed extremely weak nematicidal activity at the highest concentration (0.004%) (12.7±2.1%), and therefore had no overall development value. Fumaric acid consistently showed no significant nematicidal activity.

[0027] Example 2: LC-100 of fumaric acid and its derivatives against soybean cyst nematodes 50 Value determination To further evaluate the nematicidal activity of the compounds, regression analysis was performed with mortality rate as the dependent variable and concentration as the independent variable to calculate the median lethal concentration (LC50) of each compound against soybean cyst nematode. 50 The results are shown in Table 3.

[0028] Table 3. LC-100 of fumaric acid and its derivatives against soybean cyst nematode 50 Value test Table 3 shows that the LC50 of diethyl fumarate... 50 The value was 10.05 ± 0.26 μg / mL, which was on the same order of magnitude as the positive control fluopyram (8.52 ± 0.11 μg / mL), indicating excellent nematicidal activity. The LC50 of fumaric acid... 50 The value was as high as 684.45 ± 29.49 μg / mL, indicating extremely weak nematicidal activity, and therefore it has no development value.

[0029] Example 3: Inhibitory effect of diethyl fumarate on cyst formation of soybean cyst nematode under pot cultivation conditions To verify the line control effect of diethyl fumarate in practical applications, a pot experiment was conducted in this embodiment to determine the number of sporangia in soybean roots after treatment with the agent.

[0030] 1. Experimental Materials Test nematode: Soybean cyst nematode race 3; Soybean variety tested: Williams 82; Test reagents: Fumaric acid, Diethyl fumarate. Cultivation substrate: sterilized soil, sterilized sand, PVC pipe.

[0031] 2. Experimental Methods According to the previous LC 50 For activity data, second-instar larvae of soybean cyst nematodes were treated with 10 μg / mL fumaric acid and diethyl fumarate solutions for 48 h, respectively. Nematodes treated with water served as the control group (CK). After treatment, each treatment group was ensured to have 500 live nematodes, which were then inoculated into the root system of Williams 82 soybean plants. After incubation at 28℃ for 30 days (30 dpi), the number of sporangia on the roots of individual plants in each treatment was counted, with each treatment replicated 5 times.

[0032] 3. Experimental Results The statistical results are shown in Table 4 and Figure 1 As shown.

[0033] Table 4. Number of sporangia per soybean plant under different treatments Table 4 shows that the control group had the highest number of sporangia per plant, the fumaric acid treatment group showed a slight decrease, and the diethyl fumarate treatment group had the lowest number of sporangia. Statistical analysis indicated that the number of sporangia in the diethyl fumarate treatment group was significantly lower than that in the control group. Figure 1 ; P <0.05), while there was no significant difference between the fumaric acid treatment group and the control group. This indicates that diethyl fumarate significantly inhibited the formation of cysts by soybean cyst nematodes on soybean roots, while the inhibitory effect of fumaric acid did not reach statistical significance.

[0034] The embodiments described above are merely preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Various modifications and improvements made by those skilled in the art to the technical solutions of the present invention without departing from the spirit of the present invention should fall within the protection scope defined by the claims of the present invention.

Claims

1. Application of diethyl fumarate in the preparation of nematicides.

2. Use according to claim 1, wherein The nematicide is a soybean cyst nematicide.

3. A nematicide, characterized in that, Diethyl fumarate is the active ingredient.

4. The nematicide as described in claim 3, characterized in that, The concentration of diethyl fumarate is 0.001%-0.004%.

5. The application of diethyl fumarate or the nematicide according to claim 3 or 4 in the control of soybean cyst nematode.

6. The application as described in claim 5, characterized in that, The control of soybean cyst nematode includes killing soybean cyst nematodes and inhibiting the formation of soybean cyst nematodes.

7. The application as described in claim 5, characterized in that, The concentration of diethyl fumarate is 0.001%-0.004%.

8. A method for controlling soybean cyst nematodes, characterized in that, The step includes applying diethyl fumarate or the nematicide as described in claim 3 or 4 to the rhizosphere soil.

9. The method as described in claim 8, characterized in that, The control of soybean cyst nematode includes killing soybean cyst nematodes and inhibiting the formation of soybean cyst nematodes.

10. The method as described in claim 8, characterized in that, The concentration of diethyl fumarate is 0.001%-0.004%.

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