Composition for controlling root knot nematodes

Abstract

The present invention relates to a composition for controlling nematodes and, more specifically, to a composition for controlling root knot nematodes. The composition for controlling root knot nematodes, according to the present invention, comprises benzylidene acetone. The composition comprising benzylidene acetone for controlling root knot nematodes, according to the present invention, has an excellent nematicidal ability, and thus is environmentally friendly and can efficiently control root knot nematodes.

Description

Composition for controlling root-knot nematodes

[0001] The present invention relates to a composition for controlling nematodes, and more particularly, to a composition for controlling root-knot nematodes.

[0002] The first plant-parasitic nematode discovered worldwide was a nematode that attacked wheat in 1743. Following the discovery of the wheat nematode in Korea in 1919, root-knot nematodes, which cause significant damage to various flowers and vegetables, began to be discovered in 1977. Currently, extensive research is being conducted to control these root-knot nematodes.

[0003] Nematodes are found in all soils, and while some can be seen with the naked eye, most plant-parasitic nematodes can be observed under a microscope. Nematodes are thread-like, without segments, and move like snakes in aquatic environments. These parasitic nematodes can infest all parts of plants, including flower buds, leaves, stems, and roots. Root-based nematodes, in particular, have been reported to have a profound impact on crop growth.

[0004] In the case of root-knot nematodes, the life cycle progresses from egg to first instar larva, second instar larva, third instar larva, fourth instar larva, adult, and egg laying, taking approximately 5 to 7 weeks. Therefore, when adult root-knot nematodes are discovered on farms, the damage often progresses rapidly, and the extent of the damage can be considered extreme.

[0005] The nematodes currently occurring in Korea are causing significant damage to crops such as melons, strawberries, cucumbers, tomatoes, peppers, and cabbages that are grown in continuous cropping.

[0006] Nematodes have a very high correlation with other plant pathogens (viruses, diseases). This is because the nematodes' mouthparts damage the root mucosa, and this damage to the root mucosa often leads to secondary infections such as plant viral and bacterial diseases. The fact that nematodes transmit viruses began when it was discovered that the spear nematode transmits viral diseases to grapevines. In fact, root-knot nematodes also promote damage caused by various fungi such as Phragium, Rhizoctonia, Pythium, and Phytophthora. Therefore, the actual damage to crops caused by nematodes amounts to about 15-40% in the United States, and in reality, when secondary fungal damage is compounded during cultivation, the damage to crops caused by nematodes often reaches 60-70%.

[0007] It is known that the control methods for these nematodes include fallow, flooding, crop rotation, organic fertilizer application, cultivation of resistant plants, biological control, physical control, and chemical control, but most of these methods are difficult to apply in reality in farms, and in the case of chemical control, there is a problem that they cause destruction of the ecosystem due to soil contamination by residual toxicity due to non-selective toxicity.

[0008] As a biological control method, Korean Patent Publication No. 2007-0065938 discloses a composition for controlling root-knot nematodes containing eugenol isolated and purified from cloves, Korean Patent Registration No. 2019829 discloses a composition for controlling root-knot nematodes containing an anthraquinone compound derived from Photorhabdus temperata temprata, and Korean Patent Publication No. 2024-0001993 discloses a Boberia bassiana jef-503 strain having a root-knot nematode control effect, but there is a need to develop a nematicide having a more excellent nematicidal effect.

[0009] Accordingly, the inventors of the present invention have made efforts to develop a biological control agent for root-knot nematodes, and as a result, have confirmed that benzylideneacetone isolated from a culture solution of Xenorhabdus nematophila, a symbiotic bacterium derived from an entomopathogenic nematode, has excellent nematicidal activity against root-knot nematodes, thereby completing the present invention.

[0010] The purpose of the present invention is to provide a composition for controlling root-knot nematodes derived from natural products, which has excellent nematicidal activity without causing soil contamination or plant damage due to residual toxicity, and a nematode controlling preparation comprising the same.

[0011] To achieve the above purpose, the present invention provides a composition for controlling root-knot nematodes containing benzylideneacetone.

[0012] In the present invention, the composition for controlling root-knot nematodes is characterized in that it additionally contains garlic oil.

[0013] In the present invention, the composition for controlling root-knot nematodes is characterized by comprising 1 to 10 volume% of benzylidene acetone, 5 to 30 volume% of garlic oil, 30 to 55 volume% of emulsifier, and 25 to 55 volume% of solvent.

[0014] The present invention also provides a root-knot nematode control formulation comprising the above root-knot nematode control composition as an effective ingredient.

[0015] The composition for controlling root-knot nematodes containing benzylideneacetone according to the present invention has excellent nematicidal activity, and thus is environmentally friendly and has the effect of efficiently controlling root-knot nematodes.

[0016]

[0017] Figure 1 is a microscopic photograph evaluating the nematicidal activity against root-knot nematodes at different dilution ratios of benzylideneacetone.

[0018] Figure 2 is a microscopic photograph evaluating the nematicidal activity against root-knot nematodes according to the dilution ratio of a natural product.

[0019] Figure 3 is a microscopic photograph evaluating the nematicidal activity against root-knot nematodes according to the dilution ratio of a root-knot nematode control agent containing benzylideneacetone and garlic oil.

[0020]

[0021] In the present invention, an attempt was made to search for a substance having nematicidal activity.

[0022] In the present invention, the nematicidal activity of benzylideneacetone, plant-derived substances and plant extracts isolated from a culture solution of Xenorhabdus nematophila, a symbiotic bacterium derived from entomopathogenic nematodes, was evaluated.

[0023] That is, in one embodiment of the present invention, nematicidal activity against root-knot nematodes (Meloidogyne spp) was confirmed for benzylidene acetone, garlic oil, oregano, citronella, clover, geraniol, palmarosa, cucumber, thymol, canola oil, neem oil, derris extract, and soybean extract. As a result, it was confirmed that nematicidal activity was excellent when benzylidene acetone was used alone or when garlic oil was mixed with benzylidene acetone.

[0024] Accordingly, the present invention relates, in one aspect, to a composition for controlling root-knot nematodes comprising benzylideneacetone.

[0025] The above benzylideneacetone (trans-4-phenyl-3-buten-2-one) is a monoterpene compound that can be isolated from a culture solution of Xenorhabdus nematophila, an entomopathogenic bacterium, or can be used in a synthetic form.

[0026] The composition for controlling root-knot nematodes according to the present invention exhibits excellent nematicidal activity against root-knot nematodes even when containing benzylideneacetone alone, but may additionally contain garlic oil to exhibit a higher and more stable nematicidal rate.

[0027] A composition for controlling root-knot nematodes with added garlic oil may contain 1 to 10 volume% of benzylidene acetone, 5 to 30 volume% of garlic oil, 30 to 55 volume% of emulsifier, and 25 to 55 volume% of solvent. If the contents of garlic oil and benzylidene acetone are outside the above range, the synergistic effect may not be high.

[0028] In the present invention, the composition for controlling root-knot nematodes may be used alone, but is not particularly limited thereto, and may additionally contain an appropriate diluent or excipient depending on the formulation and / or intended use of the composition. The excipient may be a typical material depending on the formulation, and when formulated, a filler, bulking agent, wetting agent, disintegrant, or surfactant may be used. Representative diluents or excipients may include water, dextrin, calcium carbonate, lactose, propylene glycol, liquid paraffin, and physiological saline, and more preferably, double-distilled water containing 0.1 ml of Triton X-100 per liter (L) may be used.

[0029] Hereinafter, the present invention will be described in more detail through examples. These examples are intended solely to illustrate the present invention, and it will be apparent to those skilled in the art that the scope of the present invention is not limited by these examples.

[0030] Example 1: Confirmation of nematicidal activity of benzylideneacetone

[0031] The nematodes used in the experiment were Meloidogyne spp. obtained from Pusan ​​National University and propagated on tomatoes by Ecowin Co., Ltd. and used in the experiment. Tomato roots infected with root-knot nematodes were washed in running water, cut into pieces of about 1 cm, and 50 g of the cut roots were placed in a 500 ml round flask containing 200 ml of 0.5% sodium hypochloride (NaOCl). The flask was sealed with a rubber stopper and shaken vigorously for 3 minutes to sterilize the roots and burst the egg sacs. The solution was then passed through a 500-mesh overlapping 200-mesh screen and washed with distilled water to remove the NaOCl solution and recover the root-knot nematode eggs. Next, the recovered root-knot nematode eggs were surface-sterilized with 0.5% NaOCl for 10 minutes, washed thoroughly with sterile distilled water, placed in a phosphate buffer solution (1%) containing sterile saline, and hatched in an incubator at 25°C.

[0032] Benzylideneacetone was purchased from Sigma Aldrich and used.

[0033] Benzylidene acetone: emulsifier (polyoxyethylene dodecyl mono ether): auxiliary material (ethanol) were mixed in a weight ratio of 3:4:3.

[0034] After that, 50 mL of the diluted solution, in which the concentration of benzylidene acetone was diluted 100 times (3,000 ppm), 500 times (600 ppm), 1,000 times (300 ppm), 2,500 times (120 ppm), and 5,000 times (60 ppm), was added to a 250 mL Erlenmeyer flask and inoculated onto root-knot nematodes.

[0035] After inoculation with root-knot nematodes, the mixture was left in a shaking incubator at 25°C and 800 rpm for 7 days, and 1 ml was added to a 45 mm Petri dish with grid marks at 5 mm intervals every 24 hours. The number of live and dead root-knot nematodes was counted under a microscope, and the results are shown in Table 1 and Fig. 1. The control group was left untreated.

[0036] Public nematode killing rate (%) / 2 days 100 times (3,000 PPM) 500 times (600 PPM) 1000 times (300 PPM) 2500 times (120 PPM) 5000 times (60 PPM) Benzylidene acetone 100 100 100 100 100 Control 00 00

[0037] From Table 1, benzylideneacetone exhibited 100% nematicidal activity in all 100- to 5000-fold dilutions.

[0038] Example 2: Confirmation of nematicidal activity of natural products

[0039] The natural vegetable oils and extracts in Table 2 were mixed with an emulsifier (Polyoxyethylene dodecyl monoether) in a 5:5 (weight ratio). Afterwards, the concentration of the natural products was diluted 500 times (1,000 ppm) and inoculated into root-knot nematodes in the same manner as in Example 1. While leaving at room temperature, 1 ml was added to a 45 mm Petri dish with grids marked at 5 mm intervals every 24 hours, and the number of live and dead root-knot nematodes was counted under a microscope, and the results are shown in Table 2 and Fig. 2. The control group was left untreated.

[0040] Sample nematode killing rate (%) / 2 days Sample nematode killing rate (%) / 2 days Untreated 0 Polyoxyethylene dodecyl mono ether 0 Garlic oil 100 Cucumber oil 82.05 Oregano oil 79.66 Thymol 89.19 Citronella oil 84.78 Canola oil 72.22 Clover oil 100 Neem oil 70.37 Geraniol oil 100 Derris extract 92.30 Palmarosa oil 100 Sophora chinensis extract 77.19

[0041] As shown in Table 2, diluted solutions of garlic oil, clover oil, geraniol oil, and palmarosa oil all showed 100% nematicidal activity against root-knot nematodes on the second day of treatment.

[0042] Therefore, the nematicidal activity according to the dilution ratio was additionally confirmed using garlic oil, clover oil, geraniol oil, and palmarosa oil, which showed 100% nematicidal activity at a concentration of 500 times (1,000 ppm), and the results are shown in Table 3 and Fig. 2.

[0043] Announced nematode killing rate (%) / 3 days 500 times (1,000 ppm) 1,000 times (500 ppm) 2,500 times (100 ppm) 5,000 times (50 ppm) 10,000 times (25 ppm) Garlic oil 100.00 100.00 100.00 100.00 93.75 Clover oil 100.00 85.71 43.75 29.41 13.21 Geraniol oil 100.00 100.00 60.00 48.78 25.43 Palmarosa oil 100.00 100.00 56.25 47.6 121.76 Control group-----

[0044] As shown in Table 3 and Fig. 2, garlic oil showed the best nematicidal activity, and even in a 10,000-fold (25 ppm) dilution, it showed a high nematicidal activity of more than 90% after 3 days of treatment, confirming its potential as a root-knot nematode control agent.

[0045] Example 3: Confirmation of nematicidal activity of a mixture of benzylidene acetone and garlic oil.

[0046] To confirm the synergistic effect when using benzylidene acetone and garlic oil together, a prototype was manufactured with the composition shown in Table 4.

[0047] Ingredients Product A Prototype B Prototype C Prototype D Prototype E Garlic Oil 15-15305 Benzylideneacetone-55110 Emulsifier (PLE-9) 15-3015 Emulsifier (EC-1451) 5-5 Emulsifier (BKS-1) 1035501030 Ethanol 1510101010 Water 4050151930 Total 100100100100100

[0048] Afterwards, each mixed drug was diluted 1,000-fold, 2,000-fold, 5,000-fold, 10,000-fold, 15,000-fold, 20,000-fold, and 50,000-fold, and 50 mL was placed in each 250 mL Erlenmeyer flask and inoculated onto root-knot nematodes.

[0049] After inoculation with root-knot nematodes, the mixture was left in a shaking incubator at 25°C and 800 rpm for 7 days, and 1 ml was added to a 45 mm Petri dish with grid marks at 5 mm intervals every 24 hours. The number of live and dead root-knot nematodes was counted under a microscope, and the results are shown in Table 5 and Fig. 3. The control group was left untreated.

[0050] Number of insects / total number of larvae 1,000 times 2,000 times 5,000 times 10,000 times 15,000 times 20,000 times 50,000 times Prototype A 100.00100.00100.009 0.008 0.004 6.67 7.14 Prototype B 100.00100.00100.008 1.82 5 0.002 3.007 67 Prototype C 100.00100.00100.00100.009 3.33 6 0.0015 79 Prototype D 100.00100.00100.00100.009 5.008 7.83 78.43 Prototype E100.00100.00100.0094.6783.8973.3356.98 Control group0000000

[0051] From Table 5, it was found that the test products C, D and E containing both benzylidene acetone and garlic oil had a better nematicidal effect on root-knot nematodes even at a concentration diluted 10,000 times or more than the test products containing only benzylidene acetone or garlic oil.

[0052]

[0053] While specific aspects of the present invention have been described in detail above, it will be apparent to those skilled in the art that these specific descriptions merely represent preferred embodiments and are not intended to limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

[0054]

[0055] The composition for controlling root-knot nematodes containing benzylideneacetone according to the present invention has excellent nematicidal activity, and therefore can be used as an environmentally friendly root-knot nematode control agent.

Claims

1. A composition for controlling root-knot nematodes containing benzylideneacetone.

2. A composition for controlling root-knot nematodes, characterized in that it further comprises garlic oil in the first paragraph.

3. A composition for controlling root-knot nematodes, characterized in that it comprises 1 to 10 volume% of benzylidene acetone, 5 to 30 volume% of garlic oil, 30 to 55 volume% of emulsifier, and 25 to 55 volume% of solvent in the second paragraph.

4. A root-knot nematode control formulation comprising a composition for controlling root-knot nematodes according to any one of claims 1 to 3 as an active ingredient.

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