KR101032301B1 - Composition of attractant to pine wood nematode comprising camphor as an active ingredient and diagnosis kit for the detection of pine wilt disease using the composition - Google Patents

Abstract

The present invention relates to a pine ash nematode attracting composition and a pine ash nematode disease assay kit using the same. More specifically, the present invention relates to a pine ash nematode attracting composition containing a camphor as an active ingredient, a pine ash nematode disease assay kit comprising the composition, and It relates to a method for assaying the cause of death and infection by pine reptiles using the kit. The pine ash nematode attracting composition and assay kit comprising the camphor according to the present invention has an excellent effect of attracting pine ash nematodes, and has a good attracting effect even when the camphor is present at a low concentration. Simply plugging them into the pine tree can be quickly and easily tested for the cause of the pine tree nematode infection and the cause of death by simply identifying the attracted pine tree nematode.

Pine Reptile, Camper, Attracted, Black, Kit

Description

Composition of attractant to pine wood nematode comprising camphor as an active ingredient and diagnosis kit for the detection of pine wilt disease using the composition}

The present invention relates to a pine ash nematode attracting composition and a pine ash nematode disease assay kit using the same. More specifically, the present invention relates to a pine ash nematode attracting composition containing a specific ingredient as an active ingredient, a pine ash nematode disease assay kit comprising the composition. And it relates to a method for assaying the cause of death and infection by pine reptiles using the kit.

There are many types of pathogens and pests in the soil and plants, and among them, phytoparasites are known to directly or indirectly affect the growth and development of plants. Among these plant parasitic nematodes, Bursaphelenchus xylophilus is a taxonomy belonging to the leaf nematode and the pine nematode, and 49 species have been reported all over the world. In addition, a pine nematode is a nematic nematode of about 1mm in size, which is a harmful nematode that causes the tree to die by blocking the passage of moisture and nutrients in the tree tissues. Move on to infect other healthy pines. The larvae and pupa of the horned beetle grow in the pines, and when the sky cows are allegorical and escape the pines, they retain pine reptiles in their bodies, and when the fabled larvae eat the 1-2 year old shoots of healthy pines ( Pines are infected with pine reptiles. Infected pines generally fall off on the 6th day, and the leaves begin to wither from the 20th day, and after 30 days, the leaves rapidly turn red and die within a year.

Meanwhile, pine nematodes, which are caused by pine nematodes, have been occurring in many parts of the world, including Japan, Asia, the United States, the Americas, and Europe. In Japan, nearly 2.4 million m ³ of pine and algae have been killed annually since 1941. Since it first occurred in Nanjing City in 1982, China has been severely damaging about 7 million hectares of sea horse and pine forest, and in Taiwan in 1985, the special species, Yugu Song, is in danger of death. In addition, in Korea, pine trees have been infected with re-nematodes since 1988, and up to now, pine re-nematoses are occurring in 53 cities, and as they continue to spread throughout the country, not only environmental problems but also enormous economic consequences. The loss is occurring.

Therefore, in Korea, pine tree nematodes have been designated as prohibited pests under the Plant Protection Act, and various methods have been developed to control pine tree nematode diseases. Until now, various methods have been developed to control pine tree nematode diseases. There are methods such as pesticide spraying and natural methods.

The felling fumigation method is a method of killing dead insects by cutting off dead trees infected with pine ash nematodes. The fumigant can harm humans and livestock, so be careful when using it, especially with eyes, nose, mouth and skin. There is a problem that contact should be avoided, and the felling incineration method has a definite effect on the removal of pests by cutting dead trees infected with pine ash nematodes, stacking them in a large open area, and then burning them or by using a portable simple incinerator. The problem is that the timing is extremely limited. Deforestation shredding dead trees infected with pine ash nematodes and crushing them to a size of 1.5 cm or less using a sawdust or chip making machine. This method kills larvae in the wood, and this method does not have environmental pollution. There is also a low risk of forest fires. However, there is a problem in that it is easy to drop the tree containing the worms during transportation to crush the infected tree, and it takes a lot of manpower and control costs. Insecticide spraying method can prevent pine re-nematode disease by controlling the insects before planting pine, but it can prevent the spread of damage and can be applied to a large area of forest, but it can kill the insects and destroy the ecosystem. It is necessary to develop additional pesticides by increasing resistance, and there is a problem that it is not a fundamental control method in effect. In addition, the method of using a natural enemy is an ecological control means to maintain the food chain by restoring the ecosystem, there are introduced natural use, multiplication natural use, natural protection method, etc., there is a disadvantage that the control effect is inadequate.

Therefore, it is urgent to develop a method for controlling pine reptiles at a relatively low cost and a new method for effectively preventing pine infection by pine reptiles without causing adverse effects on humans and animals.

In addition, the cause of death of pine has physical causes such as root cutting by drought, air pollution, road construction, etc., and biological causes such as various insects such as nematodes, pine bark beetles, pine needles flies, and pathogens. In order to prevent the enormous forest damage caused by these causes of death, it is necessary to promptly examine the cause of tree death and take appropriate measures. However, the only way to find out that the dead pine tree is caused by the pine reel nematode is to isolate the nematode from the dead pine tree and observe it with a microscope. Also, in order to separate nematodes from dead pines, a part of the dead pines are removed with an ax, and then made into small chips within 1 cm, and the nematodes are separated by Bermann funnel method for 24 hours. One nematode should be examined under a microscope to determine whether the isolated nematode is a pine reptile. This method requires a lot of time and effort and requires special facilities such as microscopes and Berman funneling facilities, but also can only be tested by highly trained specialists who can identify and identify nematodes.

Therefore, there is a need for the development of a new test method that can be used by non-experts to quickly and accurately determine whether pines are killed by pine re-nematodes.

Therefore, while the present inventors are studying a new method to test whether pines are infected by pine nematodes and the cause of dead pines, the camphor known as a volatile substance found in pines can effectively attract pine tree nematodes. The present invention was completed by developing a kit and a method for easily and simply and quickly assaying the cause and infection of pine by nematode nematodes using the same.

Therefore, an object of the present invention is to provide a pine reticulum attracting composition comprising a camphor (camphor) as an active ingredient.

Another object of the present invention to provide a kit for assaying pine reptile nematodes comprising the composition.

Another object of the present invention to provide a method for assaying the cause of death or infection of pine by nematode nematodes using the pine nematode assay kit.

In order to achieve the object of the present invention as described above, the present invention provides a composition for attracting pine ash nematode comprising a camphor (camphor) as an active ingredient.

In one embodiment of the present invention, the camphor included in the pine ash nematode attractant composition of the present invention may be diluted with hexane to a concentration of 10 ^-3 ~ 10 ^-5 M.

In one embodiment of the present invention, the pine ash nematode attracting composition of the present invention may further include a pesticide to attract and kill the pine ash nematode.

In another aspect, the present invention provides a kit for assaying pine reptile nematodes comprising a composition for attracting pine reptile nematodes comprising the camphor (camphor) as an active ingredient.

In one embodiment of the present invention, the pine re- nematode assay kit of the present invention is a tube, a gel (gel) disposed in the lower portion of the inside of the tube, the upper portion of the gel and a camphor absorbent containing the camphor and the tube It may be to include a stopper that can be sealed.

The present invention also provides a method for assaying the cause or infection of the pine by nematode nematode comprising the step of plugging the pine re- nematode assay kit of the present invention into the pine for the purpose of the assay.

In one embodiment of the present invention, the method for assaying the cause of death or infection of pine by the pine ash nematode of the present invention is the pine ash nematode assay kit for drilling for 8 hours in the hole in the pine It may include the step of plugging in.

The pine ash nematode attracting composition and assay kit comprising the camphor according to the present invention has an excellent effect of attracting pine ash nematodes, and has a good attracting effect even when the camphor is present at a low concentration. Simply plugging them into the pine tree can be quickly and easily tested for the cause of the pine tree nematode infection and the cause of death by simply identifying the attracted pine tree nematode.

The present invention is characterized by providing a pine ash nematode attractant composition comprising a camphor (camphor) as an active ingredient.

Camphor, also called camphor, is a fatty aromatic ketone that is a white or transparent solid like beeswax. It has a molecular formula of C ₁₀ H ₁₆ O. In addition, camphor is used in the manufacture of celluloid films due to its properties of making nitrocellulose and solid solutions. It is also used as a drug-related product for excitement, cardiovascular and irritant, analgesic and preservatives. It has been found to be used. However, it was first identified in the present invention that the camphor has an effect of attracting pine reptiles.

As the active ingredient of the composition according to the present invention, the camphor may be separated from nature or synthesized by chemical synthesis. Preferably, those separated from natural plants can be used, and more preferably those separated from pine or camphor trees can be used.

In addition, the camphor included in the pine ash nematode attraction composition according to the present invention may be diluted with a suitable solvent, preferably the solvent may be hexane. In addition, the dilution may be diluted so that the concentration of the camphor is a concentration of 10 ^-3 ~ 10 ^-5 M.

On the other hand, pine nematodes are nematodes that feed on the trees while parasitic on pine or pine trees. Females are small nematodes of 0.7-1.0 mm long and males 0.6-0.8 mm long. It penetrates into the inside of the tree tissue by the bite saltwater bovine, which has an early age, and lives in the resinous part of the wood and cortex and survives by eating the parenchymal cells of pine. In addition, pine ash nematodes survive about 35 days at room temperature, and are known to become adults through 4 larvae (1st ~ 4th) larvae, and the time required for the first generation takes about 6 days at 25 ℃. It is known that the pair has a proliferative force so that after 20 days, the pair multiplies to 200,000.

However, pine trees infected with pine re-nematodes are known as “pine AIDS disease” because necrosis of oily cells around the resin tube occurs, the trees are killed due to the blockage of moisture and nutrients, and almost 100% of the trees once infected by pine re-nematodes. Also do. Since the natural enemies of the pineal nematode, which is the mediator of the pine tree nematode, have not been put to practical use, the infection of pine tree by the pine tree nematode is very likely to lead to the extinction of massive pine forests.

To date, most of the methods developed for the control of pine nematode disease have been studied mainly as a medium-borne beetle, and no direct inhibitors or attractants have been developed for biological nematode nematodes. .

Therefore, in order to effectively control pine ash nematode, it is possible to prevent pine death from pine ash nematode by rapidly diagnosing the cause of death of pine ash or increase the control efficiency for pine ash nematode disease, thereby fundamentally controlling pine ash nematode disease.

 On the other hand, pineapple nematode attracting composition comprising a camphor (camphor) of the present invention as an active ingredient has an excellent feature to attract pine ash nematode.

In addition, the pine ash nematode attracting composition of the present invention has the effect of attracting both the distributed pine ash nematode and the growth pine ash nematode.

In the above, the term “dispersed pine worms” means that the pine worms continue to breed in the woody part of the pine tree, and then, in February-May, the larvae of the pine salted larvae, which mediate the pine worm nematode, become the pupa. At this time, the pupa gathers around the pupa, and after the distributed quaternary larvae of the pine reptile invade into the body of the larvae that become adult in the pupa, the tissues of the pine tree grow through the wounds generated when the larvae eat the healthy pine. Entering and propagating and infecting is called a distributed pine tree reptile.

The above-mentioned "proliferative pine reptiles" by the medium worms when the dispersed fourth larvae enter the healthy pine, the adult lays eggs again, the eggs larva second, third larva, fourth It refers to a form of continuous growth in infected pine trees for a lifetime without being dispersed by transforming into larvae and adult larvae, and the fact that pines die is due to these propagated pine re-nematodes.

In one embodiment of the present invention, as a result of confirming the attracting effect of the pine ash nematode on the camphor which is the active ingredient of the composition according to the present invention, the camphor of the present invention is alpha-pinene (beta-pinene) and beta-pinene (β- It was shown that there is an effect of attracting pine ash nematodes, such as pinene), and has a attracting effect on not only proliferative pine ash nematodes but also distributed pine ash nematodes (see FIGS. 2 to 4).

Therefore, the present invention provides a pine ash nematode attraction composition comprising the camphor as an active ingredient.

The pine ash nematode attracting composition according to the present invention is formulated using a method known in the art to attract pine ash nematode through a rapid or sustained effect of the active ingredient after administration to a place capable of attracting pine ash nematode. Can be. The formulations may be in the form of hydrating agents, granules, powders, emulsions, sprays, smokescreens, capsules and gels.

In addition, when the composition for attracting pine ash nematode of the present invention is a formulation of emulsion, fluidity, wetting agent, microcapsules and the like, it is used by diluting with water or an appropriate solvent so that the concentration of camphor, which is an active ingredient of the composition of the present invention, is suitable for the application. It can be, if the composition for pine ash nematode attractant of the present invention is a formulation such as aerosols, fumes and toxic handouts can be applied as usual.

In addition, the pine ash nematode attracting composition according to the present invention may be used together with other pesticides, nematicides, acaricides or fungicides for the pine ash nematode attracted by the composition. These components can be formulated individually or as a mixture in a suitable carrier, or can be formulated in combination with a suitable organic solvent.

As insecticides, acaricides and nematode eradication agents which may be further included in the composition of the present invention, for example, an organic phosphate compound, a carbamate compound, a pyrethroid compound, a neoonicotinoid compound, a benzoylphenyl Urea compounds, benzoylhydrazide compounds, thiadiazine derivatives, nellytoxin derivatives, chlorinated hydrocarbon compounds, formamidine derivatives, thiourea derivatives, phenylpyrazole compounds and the like.

Therefore, when the insecticide, acaricide and nematode exterminating agent described above are additionally included in the pine ash nematode attracting composition according to the present invention, there is an effect of attracting pine ash nematodes as well as insecticidal at the same time.

On the other hand, among the various causes of pine death, especially when pines are killed by a pine reel nematode, to determine the cause of death, finely cut dead pines and soak them in water for 24 hours, and then examine the nematodes separated from the pine under a microscope. Method has been used. However, this method has a drawback that can be tested only by experts who spend a lot of time and time and can identify nematodes through a microscope.

Accordingly, the present invention provides a kit for assaying pine nematodes, which can quickly and easily test the cause of death of pines killed by pine nematodes and whether or not by pine nematodes.

The kit for assaying pine nematode nematodes according to the present invention comprises a composition for attracting pine nematode nematodes containing the camphor as an active ingredient, and can be prepared using methods known to those skilled in the art. Preferably, the kit for assaying pine reptile nematodes according to the present invention comprises: (a) a tube, (b) a gel disposed below the inside of the tube, and (c) a camphor sorbent disposed above the gel and containing a camphor. And (d) a cap capable of sealing the tube.

Specifically, in one embodiment of the present invention, by adding agar to the tube for centrifugation and hardening, and then put the filter paper (cotton ball) treated with a camper on the agar pine pine nematode Assay kit was prepared to attract, and the schematic diagram of the kit for assaying pine nematode nematodes of the present invention prepared by the above method is shown in FIG.

In addition, the inventors of the present invention prepared for the pine nematode assay kit according to the present invention comprising a composition of the present invention containing a camphor to confirm whether the effect of the pine reetrant assay kit of the present invention can actually attract pine reptile nematode As a result of inserting the kit into the tree for the purpose of assaying, it was confirmed that the effect of attracting pine reptiles was excellent.

In particular, according to one embodiment of the present invention, in order to confirm the pine tree nematode inducement effect on the kit of the present invention, after inoculating ash pieces fungus pine ash nematode in the artificial room to the pine pieces, pathogenic pine tree nematode After the inoculation, the kit for the assay was inserted into the piece of pine, the kit of the present invention containing a camphor is a attracting effect of the kit containing alpha-pinene and beta-pinene (β-pinene) It attracted more pinewood nematodes (no statistical difference), and the attracting effect of pinewood nematodes appeared to increase as the time of treatment of the attractant material increased (see FIGS. 6A and 6B).

In addition, according to one embodiment of the present invention, the pine tree nematode attracting effect on the kit of the present invention was actually investigated in the infected trees infected with pine tree nematode, and after inserting the kit of the present invention including the camper in pine After 8 hours, the pine reptiles were confirmed to be attracted, and the attracting effect was better than alpha-pinene (α-pinene) and beta-pinene (β-pinene) (see Fig. 8).

Accordingly, the present invention provides a method for quickly and accurately testing whether pines are killed or infected by pine reptiles using the kit for re-entering pine reptiles.

The assay method according to the present invention can be carried out through the step of plugging the pine re- nematode assay kit of the present invention in the pine or dead pine suspected of infection by pine re- nematode, preferably by drilling a hole in the pine It can be carried out through the step of plugging the kit according to the invention in the hole.

In addition, the assay according to the present invention can be determined by inserting the kit of the present invention into the pine for the purpose of the assay, and after a certain time, by checking the presence or absence of pine re-entry nematode attracted to the kit. That is, in the case of dead pine, if there is a pine reel nematode attracted to the kit of the present invention, it can be determined that the cause of the death is caused by the pine reel nematode. If there is a pine reel nematode attracted to the kit, it can be judged that the pine under test has been infected with pine reel nematode and therefore is likely to die.

Hereinafter, the present invention will be described in detail by way of examples. However, these examples are intended to illustrate the present invention in more detail, and the scope of the present invention is not limited to these examples.

< Example  1>

Breeding Pine reptiles  About Camphor  Attraction test

In order to confirm whether the camphor component has the effect of attracting pine reptiles, the present inventors have to examine the growth of pine reptile nematodes, that is, the dispersed fourth larvae that have entered into the pine tree, but not to adults. Attraction experiments were conducted on pine reptiles living in pines during their lifetime. For this experiment, first, the nematode Botrytis Pine reptiles grown on cinerea were isolated using a Berman funnel method, and the separated pine reptiles were diluted to 1,000 per ml. In addition, seven filter papers (8 mm) were placed on the water agar medium to confirm the induction effect of the camphor on the isolated nematodes, and the cotton ball (cotton ball) ( 1.5 cm), and six of the seven filter papers were treated for each substance (attractable substance), and the volatiles were diluted in hexane at a concentration of 10 ^-4 M and then treated. Treated only hexane as a control. Subsequently, after treating the pine ash nematode that has not fastened 1000 pine ash nematodes prepared before the cotton ball and the pine ash nematode that has been fasted for one day, the pine ash nematode after 24 hours and 48 hours was investigated at a dark condition of 24 ° C. It was. In order to examine the pine tree nematode, the 1.2 cm medium around the filter paper was removed, and half of Petri dish (60 mm) was filled with distilled water. . For more accurate investigation, the method was repeated three times, first and second times, and the average value was calculated to obtain a result. Schematic diagram of the attraction test of the above method is shown in Figure 1, 18 volatiles used in the experimental group was purchased from Sigma-Aldrich Co. The type is shown in Table 1 below, growth type pine Attraction results over time for re-nematodes are shown in Figures 2a-3b.

Volatile Materials Used in Attraction Tests
number
volatile Types of pines from which volatiles can be obtained
Molecular Weight P. massoniana
( Taiwan freight ) P. densiflora
(Shipping) P. tunbergii
(Shipping) One α- pinene o o o 136.24 2 β- pinene o o o 136.24 3 Camphene o o o 136.24 4 Limonene o o o 136.24 5 Myrcene o 136.24 6 3- Carene o o 136.24 7 Terpinolene o 136.24 8 α- Phellandrene o 136.24 9 α- Terpinene 136.24 10 γ- Terpinene 136.24 11 Decane o 142.29 12 Bornyl acetate o 196.29 13 Terpineol anhylrous o 154.25 14 Borneol o 154.25 15 Camphor o 152.24 16 Thujone o 152.24 17 P- cymene o 134.22 18 α- Humulene o 204.36

As a result, according to the first experiment according to the three repetition experiments, there was no statistically significant difference in the attracting effect of the fasting pine reptiles, but the alpha-pinene (α-pinene) after 24 hours in the day fasting pine reptiles This attraction was found, and after 48 hours alpha-pinene, beta-pinene and the camphor of the present invention were shown to be attractive. In the second experiment, alpha-pinene, beta-pinene, and camphor were superior to other volatiles regardless of fasting.

Therefore, the above results showed that the camphor component of the present invention has an excellent effect of attracting pine reptiles such as alpha-pinene and beta-pinene, and the camphor Even at a low concentration of 10 ^-4 M, the effect of attracting pine reptiles was excellent.

< Example  2>

Camper's Attraction Test on Distributed Pine Reptiles

The present inventors carried out the following experiment to determine whether the camphor component has a attracting effect on the distributed pine re- nematode. Here, the distributed pine worm nematode is when the larvae of the beetle larvae become chrysalis, the larvae of the pine reptiles gather around the pupa and enter into the body of the allegorical larvae, and then enter the tissue of the pine tree by the worms to infect the pine tree. It is called a distributed pine tree nematode. Attracting experiments on the distributed pine re- nematode were obtained by using a Berman funnel method to disperse the distributed pine re- nematode larva obtained by dissecting freshly hatched brine larvae from an infected pine tree. Re-nematode larvae were carried out in the same manner as in Example 1 except for diluting to 200 per ml and the results are shown in FIG.

As a result, alpha-pinene, beta-pinene, and the camphor of the present invention have excellent attracting effects on the dispersed pine larva larvae, as in Example 1. Appeared.

< Example  3>

Camper's Attraction Test on Growth Pine Reptiles in Artificial Chambers

<3-1> artificial chamber production

Through Examples 1 and 2 it can be seen that the camphor component is excellent in attracting pine ash nematode. Therefore, the present inventors made a kit capable of attracting and assaying pine reptiles using the camphor component, and performed the following experiment to confirm whether the kit has an effect of attracting pine reptiles. First, an artificial chamber was manufactured to perform the experiment. For this, a piece of sound pine was cut to a length of 5 cm and an average diameter of 2.5 cm, and then a hole was cut in the center of the cut surface (3-4 cm in depth and 1 in diameter). cm), after sterilizing the cut pine pieces for 15 minutes at 121 ℃ with a sterilizer, inoculated with gray mold germ which is a food of the pine ash nematode. Then, two weeks later, 1,000 birds per 1 ml of pathogenic pine reptiles were inoculated into artificial chambers and stored at 25 ° C dark conditions. The pine ash nematode with the pathogenicity was used that was obtained from the National Forest Research Institute Southern Forest Research Institute. The manufacturing process of the artificial chamber according to the above method is shown in FIG. 5.

<3-2> Kit for Pine Reptile Test including Camper

    To prepare the assay kit for attracting pine ash nematode and assaying, 2 ml centrifuge tubes were filled with 0.5% agar, and when the agar was hardened, a volatile camphor was added to the filter paper (0.5 cm x 2.5 cm). After 10 μL treatment using two methods, a cm) method and a cotton ball (0.5 cm in diameter), the tube was placed in an artificial chamber. Each treatment was carried out in three iterations, and then attracted nematodes after 24 and 48 hours at 24 ° C dark conditions. At this time, alpha-pinene and beta-pinene, which are volatile substances, were used as positive controls, respectively, and hexane was used as the negative controls. Results according to the method are shown in Figures 6a and 6b.

As a result, the attracting effect of volatiles according to the filter paper method showed little effect after 24 hours, but alpha-pinene and camphor attracted 1,089 and 1,088 nematodes after 48 hours. Beta-pinene has been shown to attract 693 nematodes. Hexane, on the other hand, had the effect of attracting less than 160 nematodes. In addition, the attracting effect of the cotton ball method was shown to have a better attracting effect than the filter paper method. In the case of alpha-pinene after 24 hours of volatile treatment, 1,129 animals were attracted and beta-pinene ( In the case of β-pinene, 905 were attracted, and in camper, more than 1,651 were attracted. Furthermore, after 48 hours, nearly 3,050 re-nematodes were attracted to the camper.

Therefore, through the above results, it can be seen that the kit for pine ash nematode assay comprising the camper of the present invention is excellent in attracting effect on pine ash nematodes, especially in the case of cotton balls, the effect of attracting pine ash nematodes is alpha-pinene (α -pinene) and beta-pinene (β-pinene) was found to be superior.

< Example  4>

Pine tree nematode In the neck  According to the invention Of kit  Attraction test

For the pine ash nematode assay kit of the present invention prepared in Example <3-2> it was examined whether it can actually attract pine ash nematode from the tree infected with pine ash nematode. For this purpose, four holes (10cm apart, 3-4cm deep, 3mm in diameter) were drilled in the side of a piece of pine, which was cut at 8 cm in length and 10 cm in diameter. 1 cm) Inserted the assay kit of the present invention including a camper here, the kit was poured by pouring 0.5% water agar into a centrifuge tube (2 ml) and a cotton ball having a diameter of 0.5 cm was placed in the center of the tube. Insert prepared was used, wherein the cotton ball was injected with camphor, α-pinene, β-pinene as an attractant, hexane was injected as a control. The kit was then inserted into a piece of pine, and then identified and identified by the number of pine reinfestation nematodes at intervals of 0.1, 1, 4, 8, 24 and 48 hours. The model of the pine ash nematode assay kit used in the present invention is shown in Figure 7, the results according to the method is shown in Table 2 and FIG.

Types and numbers of nematodes attracted to volatile attractants

Attractant
24 hours after treatment 48 hours after treatment Pine tree nematode
( Bursaphelenchus . Xylophilus ) Young pine
Nematode
( Bursaphele nchus mucronatus ) Corrosive Nematodes
( Saprophago us nematodes ) Pine tree nematode
( Bursaphelenchus . Xylophilus ) Young pine
Nematode
( Bursaphele nchus mucronatus ) Corrosive Nematodes
( Saprophago us nematodes ) Alpha-pinene 743 (mari) 6 0 238 One 0 Beta-pinene 681 0 0 518 0 0 Camper 9190 0 0 22450 0 0 Hexane 19 0 0 11 0 0

As a result, as shown in Fig. 8, after inserting the pine nematode assay kit into the infected pine tree pine needles were attracted from 8 hours, especially in the case of alpha-pinene after 24 hours of volatile treatment, 743 birds Were attracted, 681 were attracted for beta-pinene, and 9,190 were attracted for camphor. Furthermore, after 48 hours, nearly 22,450 re-nematodes were attracted to the camper.

In addition, as shown in Table 2, as a result of confirming the types of nematodes attracted by the kit according to the present invention, it was shown that only pine ash nematodes are selectively attracted to the camper. In particular, 9,190 pine nematodes were attracted in 24 hours after treatment with the kit for assaying pine nematodes, and 22,450 were attracted after 48 hours, but no similar nematode or corrosive nematodes were attracted.

Therefore, through the above results, the inventors of the present invention, when the kit of the present invention containing a camper as an active ingredient is applied to a tree suspected of being infected by the pine or pine reptiles that died due to the excellent selective attracting effect against the pine reptiles In addition, it was found that the cause of the death and the infection caused by the pine ash nematode can be quickly and accurately tested.

So far I looked at the center of the preferred embodiment for the present invention. Those skilled in the art will appreciate that the present invention can be implemented in a modified form without departing from the essential features of the present invention. Therefore, the disclosed embodiments should be considered in an illustrative rather than a restrictive sense. The scope of the present invention is shown in the claims rather than the foregoing description, and all differences within the scope will be construed as being included in the present invention.

Figure 1 is a schematic diagram showing an experiment to attract pine reptiles by the method of filter paper (cotton ball).

 FIG. 2A is a graph showing the number of attracted re-nematodes at 24 and 48 hours after treatment of volatiles in non-fasting propagated pine re- nematodes (primary experiment results), where the serial number of the X-axis is The type of each volatile substance described in Table 1 of the present invention is shown, and ⓒ represents a control treated with hexane only.

FIG. 2B is a graph showing the number of attracted re-nematodes at 24 and 48 hours after treatment of volatiles in fast-growing pine re- nematodes fasted on a daily basis (secondary test results), where the serial number of the X-axis is The type of each volatile substance described in Table 1 of the present invention is shown, ⓒ represents a control treated with hexane only.

FIG. 3A is a graph comparing the number of attracted re-nematodes at 24 and 48 hours after treatment of volatiles in non-fasting propagated pine re- nematodes (Secondary Experimental Results), where the serial number of the X-axis is The type of each volatile substance described in Table 1 of the present invention is shown, and ⓒ represents a control treated with hexane only.

Figure 3b is a graph showing the number of attracted re-nematode after 24 hours and 48 hours after treatment of volatiles in the fasting pine reptile nematode fasted per day (second experimental results), where the serial number of the X-axis is The type of each volatile substance described in Table 1 of the present invention is shown, ⓒ represents a control treated with hexane only.

Figure 4 is a graph showing the number of pine worm nematode distributed larvae attracted 24 hours after the treatment of volatiles for the dispersed pine worm nematode, wherein the serial number of the X axis is the respective volatiles described in Table 1 of the present invention It shows the kind of substance, ⓒ shows the control which treated only hexane.

Figure 5 is a schematic diagram showing the manufacturing process of the artificial chambers to test the attracting effect for the pine re-entrant nematode kit containing a camphor according to the present invention.

Figure 6a is a kit for assaying pine reptiles comprising filter papers treated with alpha-pinene (A), beta-pinene (B), camphor (C) and hexane, respectively, as volatiles. Graphs showing the attracted number of the pine ash nematode attracted after 24 hours and 48 hours after the treatment of the volatile substances using.

6b is a kit for assaying pine reptiles comprising cotton balls treated with alpha-pinene (A), beta-pinene (B), camphor (C) and hexane, respectively, as volatiles. Graphs showing the attracted number of the pine ash nematode attracted after 24 hours and 48 hours after the treatment of the volatile substances using.

Figure 7 is a diagram showing one model of the pine reptile assay kit according to the present invention.

FIG. 8 is a cotton ball volatile material using a pine nematode assay kit including cotton balls treated with alpha-pinene, beta-pinene, camphor and hexane, respectively. It is a graph showing the number of pine tree nematode attracted after 0, 1, 1, 4, 8, 24 and 48 hours after treatment.

Claims (5)

Pine ash nematode attracting composition comprising a camphor (camphor) as an active ingredient. According to claim 1, wherein the camphor pine pine nematode attracting composition, characterized in that diluted to hexane to a concentration of 10 ^-3 ~ 10 ^-5 M. The composition of claim 1, further comprising insecticides to attract and kill pine reptiles. A kit for assaying pine nematodes, which comprises a composition for attracting pine nematodes, which contains a camphor as an active ingredient. The method of claim 4, wherein the kit, tube; A gel disposed under the tube; A camphor absorbent disposed on top of the gel and containing a camphor; And Kit for pine ash nematode assay, characterized in that it comprises a stopper to seal the tube.

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