KR20150135923A - Composition for controlling nematodes comprising an extract of Waltheria species, and method for controlling nematodes using the same - Google Patents

Abstract

The present invention relates to a composition for controlling nematodes, comprising a plant extract of Waltheria sp. as an active ingredient, and a method for controlling nematodes. Specifically, the composition for controlling nematodes according to the present invention is harmless to human body due to being originated from a natural substances, and shows excellent controlling activity against Meloidogyne spp. while not causing environmental pollution, thereby being helpfully used in developing an eco-friendly natural disinfectant and producing organic agricultural products with a high value.

Description

[0001] The present invention relates to a composition for controlling nematodes, which contains plant extracts of Wasserlia genus as an active ingredient, and a method for controlling nematodes using the same,

The present invention is based on the finding that Waltheria sp.) plant extract as an active ingredient and a method for controlling nematodes thereof.

Plant parasitic nematodes are small organisms belonging to the nematode, which are present in all the soil. Most plant parasitic nematodes are very small and can only be identified on a microscope. The plant parasitic nematode is generally thread-like, thin, spindle-shaped, symmetrical, with a length of 0.25 to 0.45 mm, very small, with no nodules on the body and wrinkles on the body surface. The body wall is composed of the epidermis, the dermis, and the muscles, and constitutes the nervous system by the central nervous system and the neural rings around the esophagus. Reproduction occurs when a female adult gives birth to an egg, and it is propagated through varieties or sympathetic reproduction. The plant parasitic nematodes hatched in the egg grow after adult stage 4, survive for about 1 month and develop digestive organs and reproductive organs. In the case of females, about 50 to 600 eggs Lays. The plant parasitic nematode has a needle on the tofu, and the needle is inserted into the root tissue of the plant to puncture it, absorb the nutrients of the plant tissue, or invade into the root tissue. These plant parasitic nematodes are parasitic in all parts such as seeds, leaves, stems, and roots of plants. Depending on the species, the plant parasitic nematodes may be exogenous, And inserting the nutrients into the plant body. In addition, there are some migratory migratory migratory migratory migratory migratory migratory migratory migrants.

Nematodes are classified into bacterial nematodes, food-borne nematodes, caustic nematodes, predatory nematodes, and vegetative nematodes according to their prey. Plant nematode nematodes are one of the main causes of the sequence and cause severe damage depending on the crops. There are thousands of phytoplankton nematodes. Depending on the morphology, morphology, and host species, there are many species of Meloidogyne spp., Globodera spp., Anguina spp., Ditylenchus spp. ahyeop nematode (Aphalenchoides spp.), root rot nematode (Pratylenchus spp.), waehwa nematode (Tylenchorhychus spp.), citrus nematode (Rotylenchulus spp.) and the like.

The damage caused by plant parasitic nematodes is slight when the densities of nematodes in the soil are low, but when the density of the plants grows as the crop grows, the host plants are fatal and damaged. Plants infected with plant parasitic nematodes are less resistant to pathogens, thereby promoting or intensifying the onset of pathogens, and some nematodes mediate plant pathogenic viruses.

The root-knot nematode is one of the most damaging insect pests and is known as a host of about 700 plants. Among them, sweet potato root nematodes ( Meloidogyne incognita ), carrot root nematode ( Meloidogyne hapla ), Java rootstock nematode ( Meloidogyne javanica ) and peanut root nematode ( Meloidogyne arenaria ). It is the most difficult and hardly harmful insect pest which damages fruits, such as melon, cucumber, tomato, strawberry pepper and watermelon, and ginseng.

In order to control root nodule nematodes, fungicides such as methyl bromide and fosthizate are mainly used. However, these chemical controls have various problems such as the risk of human contraction, residual toxicity, environmental pollution, and ecosystem disturbance. In addition, the abolition of methyl bromide pesticides, which was used as an alternative to organophosphorus and carbamate pesticides, has demanded the development of safer nematicides. In recent years, more and more consumers are demanding agricultural products that are environmentally friendly. Therefore, various researches are being carried out on the development of alternative substances that are safe for man-made or environment and effective for controlling root-knot nematodes.

In order to alleviate the damage caused by plant parasitic nematodes, efforts have been made to control plant parasitic nematodes. Typical control methods include agricultural control, physical control, chemical control and biological control. Cultivar control methods include restraining, inundation treatment, rotation, adjustment of cultivation time, reforestation of resistant cultivars, and cultivation of attracted plants for a certain period of time. This kind of cultivation control is nothing but a fundamental mitigation of plant parasitic nematodes, mitigating and alleviating the damage.

As a physical control method, hot boil disinfection method, electrothermal warming method, ultraviolet ray irradiation and ultrasound method have been tried. However, in the case of hot boil disinfection method, only the temperature is elevated and the sunlight is strong, If you are not sure about the efficacy of the control by hot water sterilization law. The heat transfer method is a method of controlling plant parasitic nematodes by installing an electric heating line in the ground and warming the cultivated land using the electric heating line. This method has a disadvantage in that it is limited to the intermittent period in which the crop is not cultivated, and the facility and the maintenance cost are excessively consumed.

The present inventors intend to use the plant extract as an environmentally friendly plant-based nematode control method. Plants have produced secondary metabolites that have resisted attacks on other organisms for a long time. Therefore, it is considered that the root-knot nematodes can be effectively controlled by using these plants.

It is an object of the present invention to provide a medicament for the treatment of Waltheria sp.) plant extract as an active ingredient, and a method for controlling nematodes thereof.

In order to solve the above-mentioned object, the present invention relates to a medicament comprising Waltheria sp.) plant extract as an active ingredient.

The present invention also provides a method of controlling nematodes, which comprises treating a plant of the genus Waltheraceae to a soil, a plant or a seed of a plant.

The present invention relates to a composition for controlling nematodes containing a plant extract of Waltheria sp. As an active ingredient and a method for controlling nematodes. The composition for controlling nematodes according to the present invention comprises a nematode- It shows excellent control activity against root-knot nematodes without inducing pollution. Therefore, it can be useful for the development of environment-friendly natural product fungicide and production of high value-added organic agricultural products.

Fig. 1 shows the nematode activity of a second-instar larva of Meloidogyne incognita at 500 μg / ml level of seven Waltheria sp. Plant extracts.
Figure 2 shows the nematode activity against tomato root nematode according to various concentrations of six Waltheria sp. Plant extracts.

Hereinafter, the present invention will be described in detail.

The present invention is based on the finding that Waltheria sp.) plant extract as an active ingredient.

The Waltheria indica , Waltheria americana , Waltheria glomerata , Waltheria < RTI ID = 0.0 > calcicola , Waltheria It may be tomentosa) or wolsse La Patria help dinha (Waltheria douradinha), Ria wolsse Indica (Waltheria indica , Waltheria americana or Waltheria glomerata .

Preferably, the Wortella genus plant extract is produced by a manufacturing method comprising the following steps, but not limited thereto:

1) extracting an extract of Solanaceae with an extraction solvent;

2) filtering the extract of step 1); And

3) Concentrating the filtered extract of step 2) under reduced pressure and drying.

In this method, the Wasselia plant of step 1) may be used without limitation, such as cultivated or commercially available plants.

The extraction solvent is preferably water, alcohol or a mixture thereof. As the alcohol, it is preferable to use a C ₁ to C ₂ lower alcohol. As the lower alcohol, 30% ethanol, 50% ethanol, 70% ethanol or methanol is preferably used, and most preferably 70% good. The extraction method is preferably, but not limited to, decompression high-temperature extraction, hot water extraction, reflux extraction, hot water extraction, cold extraction, room temperature extraction, ultrasonic extraction or steam extraction. It is preferable to add 1 to 10 times the amount of the above-mentioned extraction solvent to the amount of Wasseriaca plant. The extraction temperature is preferably 30 to 100 DEG C, but is not limited thereto. The extraction time is preferably from 2 to 48 hours, but is not limited thereto. In addition, the number of times of extraction is preferably 2 to 5 times, but is not limited thereto.

In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

The nematode was identified as Meloidogyne incognita ), carrot root nematode ( Meloidogyne hapla ), Java rootstock nematode ( Meloidogyne for example, can be the genus Javanica , the peanut nematodes Meloidogyne arenaria , the Heterodera spp., the Pratylenchus spp., the Anguina spp. or the Duzenchus spp.

The composition for controlling nematode according to the present invention may further comprise, in addition to the active ingredient, a substance contained in an insecticide or a bactericide commonly used in the art.

The nematode controlling composition according to the present invention can be formulated into various formulations known in the field of agrochemicals, and any formulating method conventionally used in the field of agrochemicals can be used for formulation.

The nematocidal composition of the present invention can be formulated into a formulation preferably in the form of a wettable powder, a granule, a powder, an emulsion, a spray, a film, a capsule or a gel, and is contacted through a formulation such as a donut- It is preferable to provide it as an agent. In the present invention, it can be used in the form of a wetting agent, powder, emulsion, preferably a wetting agent .

The composition is preferably applied to crops or soil. More specifically, the composition is applied to the seeds of plants, the seeds of plants are dipped, the plants are drenched, But it is not limited to such a method, but it is preferable that the treatment is carried out by any one or more methods selected from the group consisting of spraying on plants, spraying on the foliage of plants, and treatment of floating on the plant.

The composition may be used as a general pesticide such as a plant disease controlling agent, a seed coating agent, a microbial nutrient, a soil improving agent and the like.

In a specific example of the present invention, in order to confirm the nematode activity of the methanolic extract of the genus Wasserlia to tomato root nematode, the larvae of the tomato plant root-infected tomato root were used to isolate the larvae of the second instar larva The nematode activity was examined by a 96-well microplate bioassay. After the treatment, it was observed under a microscope of magnification of 40 magnification. The presence or absence of lethality of the larvae was judged to be a dead larva (%) Were investigated. As a result, the extracts of Wasserella spp. Showed higher than 90% viable nematicidal activity at the level of 500 .mu.g / ml except for the first sample ( Waltheria douradinha leaf extract) (see FIG. 1).

The present inventors also confirmed the carnivorous activity at concentrations of less than 500 μg / ml (250, 125, 63, 32, 16 and 8 μg / ml) indica ( Waltheria indica leaf and stem extract) showed the highest carnivorous activity at 70% or higher even at a low concentration of 31 ㎍ / ml. americana Leaf and stem extract) and 4 ( Waltheria indica ( Waltheria douradinha leaf, stem and flower extract) and No. 6 ( Waltheria glomerata leaf and stem extract) samples showed 125 ㎍ / / ml and more than 70%. Almost all Waltheria indica, Waltheria americana, and Waltheria glomerata showed high nematode activity against tomato root nematode regardless of site and region, although the nematode activity was slightly different according to region, species and site (see Fig. 2 ).

Therefore, the composition for controlling nematode of the present invention can be used as an environment-friendly nematode controlling composition which is derived from a natural product and is harmless to human axis and shows excellent control activity against root-knot nematodes without causing environmental pollution.

The present invention provides a method for controlling nematodes comprising the step of treating a Wortella genus plant extract to a soil, a plant or a seed of a plant.

The < RTI ID = 0.0 > Waltheria < / RTI > indica , Waltheria americana , Waltheria glomerata , Waltheria calcicola , Waltheria It may be tomentosa) or wolsse La Patria help dinha (Waltheria douradinha), Ria wolsse Indica (Waltheria indica , Waltheria americana or Waltheria glomerata .

It is preferable that the extract is used in a liquid state by dipping or spraying on plants, seeds of plants, or coated on seeds.

Since the nematocarcinoma composition of the present invention is nontoxic to human axons and exhibits excellent control activity against nematodes without causing environmental pollution, it is useful for the development of an environmentally friendly natural disinfectant and production of high value-added organic agricultural products .

Hereinafter, the present invention will be described in detail by the following examples.

However, the following examples are illustrative of the present invention, and the scope of the present invention is not limited by the following examples.

< Example  1> Waltheria  genus( Waltheria sp .) Preparation of plant extracts

It was found that the extracts of Wasserianaceae plant extracts were highly active during the selection of excellent nematode activity for tomato root roots nematodes with various plants. To produce Methanol extract of Wasserlia plant 10 L of 70% methanol was added to 0.1 Kg of the dried sample, and the mixture was extracted at room temperature for 7 days and concentrated under reduced pressure or purchased from the plant extract bank of Korea Research Institute of Bioscience and Biotechnology. Table 1 shows the information on the seven Wasserian plant extracts used in this experiment.

number Scientific name part area One Waltheria douradinha leaf Brazil 2 Waltheria indica Leaf, stem, flower United States of America 3 Waltheria americana Leaves, stem Vietnam 4 Waltheria indica leaf Ghana 5 Waltheria indica Root Ghana 6 Waltheria glomerata Leaves, stem Costa Rica 7 Waltheria indica Leaves, stem Nigeria

Specifically, the prepared extract was dissolved in methanol, and then concentrated again with methanol to prepare a 100-fold concentrated sample, and then each sample was diluted to a final concentration of 1,000 μg / ml, 500 μg / ml, 250 μg / ml, 63 ㎍ / ml, 32 ㎍ / ml, 16 ㎍ / ml, and 8 ㎍ / ml, respectively. The untreated group was treated with 0.5 μl of methanol. After the sample was treated, the 96-well plate was stirred for 30 seconds so that the samples could be mixed well. Then, the plate was placed in a plastic container with relative humidity of 100%, sealed with a lap, and stored in an incubator (25 ± 2 ° C).

< Example  2> Tomato root-knot nematodes Meloidogyne incognita ) For Nematode  Verify Active

In order to examine the nematode activity of the tomato root roots nematode of the methanol extract of the genus Waltheria plant prepared in Example 1, the following experiment was conducted.

Specifically, the roots of the tomato plants infested with tomato root blight nematode were washed with tap water to remove foreign matter. The well-washed roots were cut into 2 cm size, placed in a blender, and 0.5% sodium hypochlorite solution was added to the roots so as to be submerged, followed by pulverization for 1 minute. After crushing, the eggs were separated by using 0.02 ㎛ sieve. The isolated eggs were used for the experiment by isolating the larvae using the funnel method (Baermann funnel). The nematode activity of the second instar larva of tomato root roots was investigated by 96-well microplate bioassay. Primary screening was performed by inoculating 49.5 ㎕ of tomato bullous nematode 2nd instar larvae in a 96-well plate at a density of 20 ~ 30 per 50 ㎕. The extract of the present invention was treated at 500 / / ml for 72 hours, and observed under a microscope at a magnification of 40. The presence or absence of lethality of the larvae was measured by shaking the well plate, The larvae were judged to be living larvae and the live nematode activity (%) was investigated. Methanol (1%) was used as a non - treated control, and abomectin (1 ㎍ / ml) was used as a control. The experiment was carried out with 3 repetitions.

As a result, as shown in Fig. 1, the extracts of Wasserella spp. Showed different nematode activities at the level of 500 ㎍ / ml, except for the sample No. 1 ( Waltheria douradinha leaf extract) (FIG. 1). In the plants treated with these extracts, the tomato root nematode was killed and the entire body was stretched out on the day, while there was no mobility. On the other hand, the control agent showed 95% strong carnivorous activity at the level of 1 ㎍ / ml.

< Example  3> at various concentrations Tomato rootstock nematode  About Nematode  Verify Active

One sample with low activity at 500 / / ml ( Waltheria douradinha Leaf Extract) was investigated in the same manner as in <Example 1> above at a concentration of 500 μg / ml or less.

As a result, FIG. 5, as shown in 2 (Waltheria indica Root extract) and 7 ( Waltheria indica Leaf and stem extracts) showed the highest carnivorous activity of 70% or more even at a low concentration of 31 ㎍ / ml. No. 3 ( Waltheria americana Leaf and stem extract) and 4 ( Waltheria indica Leaf extract) showed more than 80% viable nematode activity at the concentration of 63 ㎍ / ml, and 2 ( Waltheria douradinha Leaves, stems and flower extracts) and No. 6 ( Waltheria glomerata Leaf and stem extracts) showed more than 70% carnivorous activity at concentrations of 125 ㎍ / ml and above. Although there were slightly different nematicidal activities by region, species and site, almost all Waltheria indica , Waltheria americana And Waltheria glomerata showed high nematicidal activity against tomato root nematode regardless of region or region (FIG. 2).

Claims (5)

Wolsse in australia (Waltheria sp.) plant extract as an effective ingredient.
The method according to claim 1, wherein the Wasselia plant is Waltheria indica , Waltheria &lt; RTI ID = 0.0 &gt; americana , Waltheria glomerata , Waltheria calcicola , Waltheria tomentosa and Waltheria douradinha. &lt; RTI ID = 0.0 &gt; 8. &lt; / RTI &gt;
The nematode of claim 1, wherein the nematode is selected from the group consisting of Meloidogyne incognita ), carrot root nematode ( Meloidogyne hapla ), Java rootstock nematode ( Meloidogyne javanica ), peanut nematode ( Meloidogyne arenaria spp., Heterodera spp., Pratylenchus spp., Anguina spp., and Ditylenchus spp. Wherein the nematode inhibiting composition comprises:
A method for controlling nematode comprising treating a plant of the genus Wasserici with a soil, a plant or a seed of a plant.
[Claim 5] The method for controlling nematode according to claim 4, wherein the extract is immersed in or sprayed on plants in a liquid state, or coated on seeds.

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