Application of capillary diynone in resistance to tobacco phytophthora and phytophthora infestans

Abstract

The present invention discloses the use of capillary diynone in combating tobacco phytophthora and Phytophthora infestans, belonging to the technical field of pesticides. The present invention discovers that capillary diynone can inhibit the growth of tobacco phytophthora and Phytophthora infestans, resisting attack by tobacco phytophthora, and can be used to prepare preparations for combating tobacco phytophthora and preventing tobacco black shank. Capillary diynone can also resist attack by Phytophthora infestans and can be used to prepare preparations for combating potato late blight, thus having significant significance for combating tobacco black shank and potato late blight.

Description

Technical Field

[0001] The invention relates to application of capillary diynone in resisting tobacco phytophthora and phytophthora infestans, and belongs to the technical field of pesticides. Background Art

[0002] Capillary diol has been shown to have a significant inhibitory effect on Staphylococcus aureus in vitro. It also has some inhibitory effects on Shigella dysenteriae, hemolytic Streptococcus, Pneumococcus, Corynebacterium diphtheriae, bovine and human tuberculosis, Escherichia coli, Salmonella typhi, Pseudomonas aeruginosa, Bacillus subtilis, pathogenic filaments, and dermatophytes such as Aspergillus flavus and Aspergillus versicolor. Furthermore, patents disclose that a combination of capillary diol and boscalid can control gray mold on crops.

[0003] Potato late blight has occurred in my country for many years. It is caused by infection with the pathogenic fungus Phytophthora infestans. The early symptoms of potato late blight are not obvious, with only small gray spots. It enters the middle and late stages after a few days of infection. "The fields look like they have been burned by fire, and the leaves are charred; due to the inability to photosynthesize, the potato stems and leaves soon wither and die."

[0004] Tobacco black shank, also known as tobacco black root or black stem madness, is a major soil-borne fungal disease of tobacco caused by Phytophthora nicotianae. It primarily infects the roots and base of the stems, forming dark, sunken lesions, hence the name "black shank."

[0005] The present invention firstly discovers that capillary diynone has a good inhibitory effect against tobacco phytophthora and phytophthora infestans. Summary of the Invention

[0006] The first object of the present invention is to provide an application of capillary diynone in inhibiting the growth of tobacco phytophthora mycelium, thereby providing a new prevention and treatment method for tobacco black shank disease.

[0007] The second object of the present invention is to provide the use of capillary diynone in the preparation of a preparation for inhibiting tobacco phytophthora.

[0008] The third object of the present invention is to provide the use of capilladiynone in inhibiting the growth of Phytophthora infestans hyphae.

[0009] The fourth object of the present invention is to provide the use of capillary diynone in the preparation of a preparation for inhibiting Phytophthora infestans, thereby providing a new prevention and control method for potato late blight.

[0010] Preferably, the capillary diynone of the present invention is used at a concentration of 1 to 5 mg / L.

[0011] Preferably, the preparation of the present invention is a fertilizer or a pesticide.

[0012] The capillary diynyl ketone of the present invention is homemade, commercially available or obtained in any other way.

[0013] The invention has the beneficial effects of effectively preventing and controlling tobacco black shank disease and potato late blight in agricultural production, ensuring yield and reducing economic losses. BRIEF DESCRIPTION OF THE DRAWINGS

[0014] Figure 1 This is a diagram showing the antibacterial effect of capillarin diacetyl ketone on Phytophthora infestans. DETAILED DESCRIPTION

[0015] The present invention will be further described in detail below with reference to specific embodiments, but the protection scope of the present invention is not limited to the contents described above.

[0016] Example 1

[0017] The capillary diols used in the embodiments of the present invention are synthesized by myself. In practice, capillary diols obtained by any means can be used in the present invention. The specific synthesis route is as follows:

[0018]

[0019] Synthesis of 1b: To a 50 mL round-bottom flask were added 1-phenyl-2-propyn-1-ol (1 mmol, 1a), catalyst AgNO3 (0.1 mmol), N-bromosuccinimide (NBS) (2 mmol), and 6 mL of acetone. The mixture was stirred at room temperature for 4 h, resulting in the formation of a white solid. After completion of the reaction, the solid was removed by filtration, and the filtrate was concentrated under vacuum and purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 20:1) to afford compound 1b in 80% yield.

[0020] Synthesis of 1c: To a 50-mL round-bottom flask were added hydroxylamine hydrochloride (1 mmol), piperidine (0.2 mL), the catalyst CuCl (0.02 mmol), and methanol (3 mL) in sequence, and stirred in an ice bath. Propylene (1 mmol, 1 mol / L in THF) was added at 0°C, stirred for 10 minutes, and then compound 1b (1 mmol) was added dropwise. The reaction mixture was stirred at 0°C for 1 hour. After the reaction was complete, the methanol was removed under vacuum by thin-layer chromatography (petroleum ether:ethyl acetate = 15:1, KMnO4 chromogen). The mixture was diluted with water and extracted with ethyl acetate. The organic layer was dried over anhydrous Na2SO4, concentrated under vacuum, and purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 20:1) to obtain compound 1c in a 20% yield.

[0021] Synthesis of 1d: Compound 1c (1 mmol) was dissolved in 6 mL of dichloromethane, MnO2 (10 mmol) was added, and the mixture was stirred at 25°C for 12 h. After completion of the reaction as monitored by thin-layer chromatography, MnO2 was removed by filtration using celite. The mixture was concentrated under vacuum and purified by silica gel column chromatography (eluent: petroleum ether:ethyl acetate = 10:1) to obtain compound 1d in a 75% yield.

[0022] The specific process of the antibacterial test of capillaris diacetyl ketone is as follows:

[0023] The synthesized capillarin diacetyl ketone was placed in a brown bottle and stored in a refrigerator at 4°C. Before the experiment, 0.005 g of capillarin diacetyl ketone was weighed and added to 995 μL of dimethyl sulfoxide (DMSO) solution to prepare a 5 mg / mL mother liquor. The prepared mother liquor needed to be stored in a refrigerator at 4°C.

[0024] Preparation of rye culture medium: Use an electronic balance to weigh 60g rye, 20g white sugar, 15g agar, 1.4g CaCO3, and prepare 1000ml of distilled water at the same time; first soak the rye in water for 24h, then use a wall breaker to break and filter the rye, add white sugar, agar and CaCO3 to the filtered liquid, and heat and stir until completely mixed during the addition process; after mixing, use a measuring cylinder to measure 50ml of culture medium and add it to the prepared conical flask, use a measuring cylinder to measure to ensure that the amount of culture medium in the conical flask is 50ml, and seal the conical flask with culture medium with sealing film to prevent contamination; after sealing the sealing film, you need to add another layer of newspaper on the outside to cover the sealing film, and finally put the sealed conical flask and culture dishes wrapped with newspaper and other tools needed into a high-pressure steam sterilizer for sterilization.

[0025] Preparation of poisonous plates: The prepared mother liquor of capillaris diacetyl ketone, which has been refrigerated in the refrigerator for standby use, is placed in an oven at 60°C for half an hour, and then placed in the refrigerator for cooling. After cooling, it is placed in a clean bench, and the sterilized culture medium and other tools needed are also placed in the clean bench (the clean bench needs to turn on the UV lamp and fan for treatment half an hour in advance, and the table of the clean bench should be wiped clean with alcohol). Then, 100 μL of the mother liquor of capillaris diacetyl ketone is drawn into the sterilized culture medium using a pipette, and the mother liquor of capillaris diacetyl ketone and the culture medium are fully mixed, so that the final concentration of the culture medium is 10 μg / mL. Then, it is poured into a sterilized and dried culture dish, and placed on one side of the clean bench for cooling. After cooling, the poisonous plate is obtained.

[0026] Inoculation of strains: Place the activated cultured strains into the clean bench and use a sterilized puncher to punch holes. The punching position should be the outer edge of the mycelium growth, because the mycelium at the edge has not yet aged, has good activity, and is easier to culture and grow. It is also necessary to ensure that there is mycelium growth on each cake. Then use sterilized tweezers to place the cake into the center of each culture dish. Seal the edges of the culture dish inoculated with the strain with sealing glue to prevent contamination. At the same time, write labels with a pen and place it upside down in an incubator at 18°C for culture.

[0027] Setting up the control group: An equal amount of dimethyl sulfoxide aqueous solution needs to be added to the control group, and three replicates should be set up.

[0028] Recording and processing of antibacterial results: The cross-cross method was used to measure the growth diameter of the colony on the culture dish and calculate the antibacterial rate. The antibacterial experiment was terminated when the hyphae of tobacco phytophthora or Phytophthora infestans on the control group (CK) plate were fully grown.

[0029] Colony pure tensor = colony average diameter - cake diameter (mm)

[0030] Inhibition rate (%) = [(pure growth of control colonies - pure growth of treated colonies) / pure growth of control colonies] × 100% Figure 1 It can be seen that when the concentration of capillary diynone is 10 μg / mL, the growth of mycelia of the two strains of Phytophthora infestans can be completely inhibited, and the culture time is 7 days.

[0031] According to the experimental results, when the final concentration of capillary dioxin in the culture medium is 10 μg / mL, it has an antibacterial effect on both Phytophthora fungi, but at the same concentration, the antibacterial effect of capillary dioxin on Phytophthora infestans is significantly better than that on Phytophthora nicotianae.

[0032] This example further studied the effective medium concentration of capillary diynone against Phytophthora infestans, and set a concentration gradient of 0, 1, 2, 3, 4, and 5 μg / mL, as shown in Tables 1 and 2.

[0033] Table 2 The results of the inhibition of capillary diacetyl ketone on Phytophthora infestans

[0034]

[0035] Table 2 Effective concentration of capillarin diacetyl ketone against Phytophthora infestans

[0036]

[0037] According to the experimental results in Tables 1 and 2, the inhibitory effect decreases with the increase of time. The table below shows that the effective median concentration of capillarin diacetyl ketone against Phytophthora infestans is between 1.979–3.818 μg / mL. At the same time, according to the experimental results, the inhibition rate gradually weakens with the increase of culture time, and the higher the concentration, the better the inhibitory effect.

Claims

1. Application of capillarin diacetyl ketone in inhibiting the growth of mycelium of Phytophthora infestans.

2. Application of capillaris diynone in the preparation of preparations for inhibiting Phytophthora infestans.

3. The use according to any one of claims 1 or 2, characterized in that: The usage concentration of capillarin is 1-5 mg / L.

4. The application according to claim 2, characterized in that: The preparation is a fertilizer or a pesticide.

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