Application of towel gourd extract in prevention and treatment of muskmelon gummy stem blight

Abstract

The invention discloses application of a towel gourd extract in prevention and treatment of muskmelon gummy stem blight. A preparation method of the towel gourd extract comprises the steps of towel gourd raw material treatment, organic solvent extraction and concentration. The towel gourd extract can significantly inhibit the activity of muskmelon gummy stem blight germs, is a plant extract with good biocontrol effect and no pollution, and has wide development and application prospects.

Description

Technical Field

[0001] The invention relates to the technical field of agricultural planting disease prevention and control, and in particular to application of a loofah extract in preventing and controlling melon vine blight. Background Art

[0002] Melon vine blight is caused by Stagonosporopsiscucurbitacearum Diseases that occur on melons due to infection. Melon vine blight generally harms the main vine and side vines, and sometimes petioles and leaves. In the early stage of leaf damage, yellow-brown "V"-shaped spots with unclear rings appear on the leaf margins, and then the entire leaf dies. In the early stage of petiole damage, yellow-brown oval to stripe-shaped spots appear, and then the diseased part gradually shrinks, and the branches and leaves above the diseased part die. Melon vine blight is a devastating fungal disease that harms the melon industry worldwide. It is highly contagious, develops quickly, and causes serious damage. Once infected, it will have a devastating impact on the melon industry.

[0003] Loofah( Luffa aegyptiaca Miller), a crop of the genus luffa of the Cucurbitaceae family, also known as Shenggua, Caigua, and Shuigua, is a common edible melon vegetable. Luffa contains rich nutrients and has anti-inflammatory, antioxidant, immunomodulatory and anti-cancer functions. In addition, studies have found that luffa is an effective anti-inflammatory drug with biological activity, but most of the research on luffa extracts is focused on beauty and detergents. In terms of plant pathogens, luffa ethanol extract has an antibacterial effect on apple anthracnose, Penicillium, apple ring-shaped fungus and cotton wilt, but there is no report on the antibacterial effect on melon pathogens.

[0004] In the prior art, the inhibitory base against plant pathogens is mainly spraying pesticides. Pesticides are highly targeted and will not cause harm to non-target organisms, but they have obvious problems, such as long-term harm to the soil, ecological environment, and human and animal health. For example, patent CN201310285606.X discloses a granular fertilizer containing boscalid and its application. The fertilizer, with its total mass as 100%, includes the following components in mass percentage: 0.1%-20% of pesticide active ingredient, 70%-99% of fertilizer carrier, 0.05%-9% of organic solvent, 0.05%-0.5% of surfactant, and 0.05%-0.5% of coating agent. The pesticide active ingredient here is boscalid. The combined application of boscalid and fertilizer has a good field effect on soil-borne diseases such as vine blight, damping-off, and wilt. However, the boscalid and organic solvent in the granular fertilizer disclosed in the patent will bring relatively long-term harm to crop soil, ecological environment, etc.

[0005] Patent CN202311658214.3 discloses a microbial agent for preventing and treating melon vine blight and its preparation method, wherein the microbial agent contains 10-15 parts of biological composite bacteria, 20-30 parts of diatomaceous earth, 40-50 parts of seaweed polysaccharide, 3-5 parts of zinc sulfate, 0.5-1 parts of copper sulfate, 1-3 parts of sodium molybdate, 0.2-0.5 parts of ferrous sulfate, 1-2 parts of potassium sulfate, and 0.05-0.1 parts of magnesium sulfate; wherein the biological composite bacteria include non-hygroscopic Streptomyces, Bacillus amyloliquefaciens, Trichoderma harzianum, and Pseudomonas fluorescens. The agent has a good effect on preventing and treating melon vine blight and has a good inhibitory effect on vine blight pathogens. The biological composite bacteria disclosed in the patent for inhibiting melon vine blight are microbial components, which are generally present in the field during application. The residual pesticides in the field or after the application of pesticides affect the activity of the biological composite bacteria, and the final antibacterial effect is greatly reduced.

[0006] Therefore, it is of great significance to develop new biological pesticides that inhibit melon vine blight, as the existing use of pesticides to treat melon vine blight can easily affect crop soil and ecological environment, and the existing microbial agents can be easily affected by residual pesticides in the field. Summary of the invention

[0007] The purpose of the present invention is to provide an application of a loofah extract in preventing and treating melon vine blight, so as to solve the problems that existing pesticides are easy to affect crop soil and ecological environment, and microbial agents are easily affected by residual pesticides in the field.

[0008] The purpose of the present invention is achieved through the following technical solutions: Use of a loofah extract in preparing a preparation for preventing and treating melon vine blight.

[0009] Furthermore, the preparation method of the sponge gourd extract comprises: sponge gourd raw material processing, organic solvent extraction, and concentration. Furthermore, the preparation method of the sponge gourd extract comprises: (1) Sponge gourd raw material processing: fresh and tender sponge gourd fruit is washed with water, dried and crushed, and sieved through 40 meshes to obtain sponge gourd powder; (2) Organic solvent extraction: adding organic solvent to the sponge gourd powder for dipping, and filtering after dipping; (3) Concentrating: The filtrate after filtering in step (2) is evaporated and concentrated to prepare the sponge gourd extract.

[0010] Furthermore, the organic solvent used in step (2) is 90-98% ethanol, preferably 95% ethanol, wherein 95% ethanol is 95% ethanol aqueous solution, and 90-98% has the same meaning.

[0011] Furthermore, the soaking method in step (2) is: adding 10 times the volume of ethanol to the loofah powder and fully soaking it, shaking it at a constant temperature of 20-30° C. for 8-24 hours, and filtering to obtain a filtrate and a filter residue.

[0012] Furthermore, the number of immersions is 3 times, and the specific immersion methods are as follows: first immersion: add 10 times the volume of ethanol to the loofah powder to fully immerse it, oscillate at a constant temperature of 30°C for 24 hours, and filter to obtain the first filtrate and the first filter residue; second immersion: add 5-10 times the volume of ethanol to the first filter residue to fully immerse it, oscillate at a constant temperature of 30°C for 12 hours, and filter to obtain the second filtrate and the second filter residue; third immersion: add 5-10 times the volume of ethanol to the second filter residue to fully immerse it, oscillate at a constant temperature of 30°C for 8 hours, and filter to obtain the third filtrate and the third filter residue.

[0013] Furthermore, the temperature of evaporation and concentration in step (3) is 45-55°C.

[0014] A method for preventing and controlling melon vine blight comprises contacting melon leaves with a loofah extract for prevention and control.

[0015] The beneficial effects of the present invention are: 1. The invention provides an application of a sponge gourd extract in preventing and treating melon vine blight. The sponge gourd extract can inhibit the activity of melon vine blight pathogen, effectively prevent and treat melon vine blight caused by melon vine blight pathogen, and provide a new idea for the subsequent development of biopesticides using sponge gourd extract to prevent and treat melon vine blight.

[0016] 2. The invention provides an application of a sponge gourd extract in preventing and treating melon vine blight. The sponge gourd extract is applied to inhibit the activity of melon vine blight pathogens. The sponge gourd extract is a plant secondary metabolite and can participate in natural material and energy metabolism. When applied to plants or the environment, it will not cause biological enrichment, environmental pollution, or residue.

[0017] 3. The invention provides an application of a sponge gourd extract in preventing and treating melon vine blight. The extract is prepared using sponge gourd as raw material. The raw material source is wide, the price is low, the production cost is low, the production process is simple, and it is suitable for production of various scales. BRIEF DESCRIPTION OF THE DRAWINGS

[0018] Figure 1 This is a diagram showing the antibacterial effect of the loofah extract of Example 2 of the present invention on melon vine blight bacteria; Figure 2 This is a diagram showing the effect of the sponge gourd extract of Example 3 of the present invention on preventing and treating melon vine blight. DETAILED DESCRIPTION

[0019] Definitions of terms used in the present invention: Unless otherwise stated, the initial definitions provided for the terms in this document apply to the terms throughout the specification; for terms that are not specifically defined in this document, the meaning that a person skilled in the art would give them should be given based on the disclosure and context.

[0020] The reagents and equipment used in the present invention are all known products or obtained by purchasing commercially available products.

[0021] The pathogens used in the following examples are described as follows: Stagonosporopsiscucurbitacearum , numbered 14mk020, the document source is "QTL positioning analysis of melon vine blight resistance based on SLAF_seq and BSA technology", which is preserved and provided by the Vegetable Institute of Ningbo Academy of Agricultural Sciences. The public can contact the unit to obtain the corresponding pathogens. The biological material is only used to repeat the relevant experiments of the present invention and cannot be used for other purposes.

[0022] In order to more clearly illustrate the technical solution of the present invention, the specific implementation methods are described as follows: Embodiment 1 Preparation of Luffa Extract (1) Wash the fresh loofah fruit with water, dry it, and then crush it into powder, and sieve it through a 40-mesh sieve to obtain loofah powder.

[0023] (2) The extraction was performed by three immersion methods. Specifically, 50 g of loofah powder was weighed and placed in a sealed conical flask, and then fully immersed in 10 times the volume of 95% ethanol, and oscillated at a constant temperature of 30°C and 120 r / min for 24 h, and then filtered under reduced pressure to obtain the first filtrate and the first filter residue; the first filter residue was further immersed in 6 times the volume of 95% ethanol, and oscillated at a constant temperature of 30°C and 120 r / min for 12 h, and then filtered under reduced pressure to obtain the second filtrate and the second filter residue; the second filter residue was further immersed in 6 times the volume of 95% ethanol, and oscillated at a constant temperature of 30°C and 120 r / min for 8 h, and then filtered under reduced pressure at 50°C to obtain the third filtrate and the third filter residue.

[0024] (3) The three filtrates obtained by the three immersion methods in step (2), namely the first filtrate, the second filtrate and the third filtrate, are combined by rotary evaporation and concentrated by evaporation at 55° C. to obtain a loofah extract with a concentration of 1 g / mL. Here, 1 g / mL of loofah extract means that 1 ml of loofah extract contains 1 g of loofah sample substance. Here, the loofah sample substance is defined as the fresh loofah fruit before washing in the above step (1).

[0025] Embodiment 2 Determination of antibacterial activity of sponge gourd extract against melon vine blight pathogen The growth rate method was used to determine the antibacterial activity of melon vine blight pathogen.

[0026] Preparation of the verification solution of the sponge gourd extract: using distilled water as the solvent, dissolve the 1 g / mL sponge gourd extract prepared in Example 1 to a final concentration of 10% sponge gourd extract (0.1 g / mL) for testing, where 0.1 g / mL means that 1 ml of sponge gourd extract contains 1 g of sponge gourd sample material.

[0027] The prepared sponge gourd extract was sterilized and filtered through a 0.22 μm microporous filter membrane, and the PDA medium was heated and melted and then cooled to about 50°C, and poured into a culture plate, with 15 ml per dish. Different volumes (1000 μl, 750 μl, 640 μl, 500 μl, 250 μl) of sponge gourd extract verification solution were added and mixed evenly, so that the final concentrations of sponge gourd extract in each culture dish were: 6.25 mg / ml, 4.76 mg / ml, 4.10 mg / ml, 3.23 mg / ml, 1.64 mg / ml, and after solidification, the melon vine blight pathogens expanded on PDA were cut with a 6 mm diameter puncher and transferred to the center of the culture medium with sponge gourd extract. Each concentration treatment was repeated for 3 dishes, and the melon vine blight pathogens cultured on the culture medium with sterile distilled water were used as the control.

[0028] The culture was placed at 25°C and the colony diameter was measured on the 6th day using the ruler cross method to obtain the inhibition rate of different concentrations of loofah extract on the growth of melon vine blight pathogen. The inhibition rate calculation formula is: inhibition rate (%) = (colony diameter of control group - colony diameter of treatment group) / colony diameter of treatment group × 100%. The specific inhibition effect of loofah extract on melon vine blight pathogen is as follows: Figure 1 The specific results of the inhibition rate of melon vine blight pathogen growth are shown in Table 1 below, where Figure 1 CK in Table 1 means the control experimental group.

[0029] Table 1 Inhibition of different concentrations of loofah extract on the growth of melon vine blight pathogen Mass concentration of loofah extract (mg / ml) Colony diameter (cm) Antibacterial rate (%) 6.25 0.80±0.02 90.70±0.23 4.76 2.19±0.21 74.50±2.40 4.10 4.42±0.27 48.64±3.16 3.23 5.27±0.48 38.76±5.52 1.64 7.70±0.00 10.47±0.00 0 (CK) 8.60 / According to the results in Table 1, the mass concentration of loofah extract reaching 6.25 mg / ml can achieve an antibacterial effect of 90.70%.

[0030] Embodiment 3 To test the efficacy of loofah extract on melon vine blight.

[0031] The in vitro leaf method was used to detect the control effect of loofah extract on melon vine blight. The melon variety was Oriental Mi No. 1, which is a common melon variety on the market. Place absorbent paper in a disposable culture dish and spray water to soak it. Pick the second true leaf when the melon has three leaves and one heart, and place the true leaf on the absorbent paper in the culture dish. Inoculate a 6mm mycelium block of vine blight fungi in the center of the leaf. Use a pipette to draw 300μl of the loofah extract verification solution prepared in Example 2 (that is, 10% loofah extract) and drop it on the mycelium block. Add clean water as a control. Each treatment was repeated 3 times. Cover the treated culture dish with a lid and seal it with a sealing film to keep it moist. Place it in an incubator at 25°C. After 72 hours, the diameter of the vine blight lesions is counted. The specific control effect of loofah extract on melon vine blight is shown in the figure below. Figure 2 The specific results of the lesion diameter of the vine blight are shown in Table 2. Figure 2 CK in Table 2 means control group.

[0032] Table 2 Control effect of loofah extract on melon vine blight Processing Name Diameter of lesion (cm) 10% Luffa Extract 0.49±0.10 CK 4.50±0.30 According to Table 2 and Figure 2 The results showed that 10% loofah extract had a significant preventive and therapeutic effect on melon vine blight.

Claims

1. Use of loofah extract in preparing a preparation for preventing and treating melon vine blight.

2. Use of the sponge gourd extract according to claim 1 in preparing a biological pesticide for preventing and treating melon vine blight, characterized in that: The preparation method of the sponge gourd extract comprises: sponge gourd raw material processing, organic solvent extraction and concentration.

3. Use of the sponge gourd extract according to claim 2 in the preparation of a biological pesticide for preventing and treating melon vine blight, characterized in that: The preparation method of the sponge gourd extract comprises: (1) Sponge gourd raw material processing: fresh and tender sponge gourd fruit is washed with water, dried and crushed, and sieved through 40 meshes to obtain sponge gourd powder; (2) Organic solvent extraction: adding organic solvent to the sponge gourd powder for dipping, and filtering after dipping; (3) Concentration: The filtrate after filtering in step (2) is evaporated and concentrated to prepare the sponge gourd extract.

4. Use of the sponge gourd extract according to claim 3 in preparing a biological pesticide for preventing and treating melon vine blight, characterized in that: The organic solvent used in step (2) is 90-98% ethanol.

5. Use of the sponge gourd extract according to claim 3 in preparing a biological pesticide for preventing and treating melon vine blight, characterized in that: The soaking method in step (2) is as follows: add ethanol to the loofah powder and fully soak it, oscillate at a constant temperature of 20-30° C. for 8-24 hours, and filter to obtain a filtrate and a filter residue.

6. Use of the sponge gourd extract according to claim 5 in the preparation of a biological pesticide for preventing and controlling melon vine blight, characterized in that: The number of immersions is 3 times; The specific impregnation method is as follows: first impregnation: add 10 times the volume of ethanol to the loofah powder to fully soak it, oscillate at a constant temperature of 30°C for 24 hours, and filter to obtain the first filtrate and the first filter residue; second impregnation: add 5-10 times the volume of ethanol to the first filter residue to fully soak it, oscillate at a constant temperature of 30°C for 12 hours, and filter to obtain the second filtrate and the second filter residue; third impregnation: add 5-10 times the volume of ethanol to the second filter residue to fully soak it, oscillate at a constant temperature of 30°C for 8 hours, and filter to obtain the third filtrate and the third filter residue.

7. Use of the sponge gourd extract according to claim 5 in preparing a biological pesticide for preventing and controlling melon vine blight, characterized in that: The temperature of evaporation and concentration in step (3) is 45-55°C.

8. A method for preventing and treating melon vine blight, characterized in that: This includes contacting the leaves of melon with a loofah extract for control.

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