JPS5924125B2 - Pine tree blight prevention method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a method for preventing powder blight, and more specifically, the present invention relates to a method for preventing powder blight.
Enchus 11gn1colus) - Arthrobotrys sp. TKBC1
This invention relates to a method for preventing blight of powders, which is characterized by applying 015-Feikokenkyori No. 5648- to the powders.

As is well known, plugs in our country - red pine, black pine,
The loss of pines such as Ryukyu pine has been increasing rapidly in recent years, and the amount of damaged timber was 15 in 1973.
The amount of damage has reached 1,000,000 cubic meters, and the area affected is also expanding, and has now spread throughout the country except for Nagano, parts of the Tohoku region, and Hokkaido.

It was in 1972 that it was discovered that the cause of this abnormal pine blight phenomenon was infection with a nematode called the pine nematode, which is less than an hour long.
The following year, it was discovered that the pine beetle was the carrier of the pine nematode.

The mechanism of pine blight caused by the pine tree nematode is as follows.

In the fall, the pine beetle chooses branches and trunks of pine trees that have withered away due to the pine tree nematode, and lays eggs there.

Inside the tree, pine tree nematodes are rapidly multiplying.

In spring, the pine beetle becomes waxy inside the tree in a chamber called a pupa, which becomes a pupa.

At that time, the pine-nosed nematodes concentrate around the cage.

This phenomenon is called the octopus effect, and is said to be caused by some kind of attractant produced by the beetle.

Immediately after the pine beetle emerges in the dance chamber, the pine beetle nematode climbs up the protrusions on the chamber wall and the hyphae of black mold, attaches itself to the body of the pine beetle, and enters the trachea through the spiracle.

Finally, an average of 1 per pine beetle
5,000 pine tree nematodes will migrate.

Adult longhorn beetles with their bodies filled with pine beetles escape from dead trees and feed on the young branches of healthy trees.

At this time, the pine tree nematodes crawl out of the spiracles and gather near the tail of the longhorn beetle, and when they touch the afterfeeding part, they attach themselves to the tree body and kill the tree.

This tree is used by the Japanese longhorn beetle as a spawning target tree in the fall.

In this way, the life cycle repeats itself. Of course, various prevention methods have been adopted at great expense to prevent blight, but it is well known that there is still no definitive prevention method and it has become a major social and economic problem. That's exactly what it says.

In other words, conventional prevention methods include: ■ Spraying preventative chemicals on healthy trees by helicopter or from the ground; ■
Methods such as cutting down damaged trees, ■ Attracting and killing the carrier pine beetle using attractants, ■ Injecting chemicals into tree trunks, and ■ Resistance breeding methods are being implemented.

However, method (■) kills longhorn beetles with chemicals during after-feeding, and before the long-horned beetles come in contact with the chemical on young branches and die, pine wood nematodes can enter the tree from the after-fed parts and kill the tree. It cannot be said to be a complete method to prevent withering because it is stored away.

In addition, since strictness is required in terms of the amount of the drug, the timing, frequency, and method of spraying, inappropriate spraying will result in no medicinal efficacy.

There is also a big concern about drug damage. Method (2) involves cutting down dead trees and killing the pine beetle larvae by peeling, burning, or spraying with chemicals, but it is impossible to cut down a large amount of dead trees in a timely manner. be.

Incineration also poses the risk of wildfires in the mountains.

Furthermore, chemical spraying is not effective against larvae and pupae that have penetrated deep into the log, and even larvae that have just hatched are ineffective unless the chemical is applied to the back side of the log.

Regarding the method (2), no substance has been developed so far that has more attractivity than dead wood, and no method has been developed to completely trap or kill the attracted longhorn beetles.

The chemicals used in method (2) are all powerful drugs that are not registered as pesticides, so it is not a method that anyone can use anywhere.

With method (2), it is not clear what causes the resistance, and the current situation is that the plant is simply propagated by grafting the remaining natural water that has escaped damage.

Moreover, the pine nematode used for the first resistance test was isolated in the Hikari region, where it is said to have moderate harmfulness, and it was isolated in the Ibaraki region, where it is said to have strong harmfulness. The secondary test using the pine tree nematode for this strain was 5.
It is scheduled to be carried out after 5 years.

Therefore, it will still take time for resistant varieties to be developed.

As mentioned above, among the conventionally practiced prevention methods (1) to (3), only methods (2) and (3) are said to have some degree of effectiveness.

However, methods ■ and ■ are only methods for exterminating the pine beetle, and are not a fundamental extermination method for the culprit, the pine nematode, which is the reason why the pine blight damage not only cannot be eradicated, but also has a tendency to spread. It can be said.

In view of the above-mentioned current situation, the present inventor is conducting systematic research from the viewpoint of preventing pine blight by fundamentally exterminating pine tree nematode.

Then, the present inventors focused on nematode-predating bacteria, which are natural enemies of nematodes.

In other words, nematode-predating fungi form special organs on the hyphae,
This name is given to an ecological group that has predatory properties that trap and kill nematodes and absorb their body contents.

Most of them belong to Deuteromycetes, and Dactylaria (Da
ctylar ia), Dactylela
lla), Monacrosporium (Monacrosp
orium) etc. are the main genera.

To date, approximately 150 species of nematode-predating fungi are known.

Research on biological control of plant-parasitic nematodes has been conducted overseas because they are natural enemies of nematodes, but there are few examples of research in Japan.

The nematode control effect that has been recognized so far is Monacrosporium ellipsosporum (Monacrosporium nematode) in a pineapple orchard in Hawaii.
There is a research report that shows that Dactylella ellipsosporum can be controlled by applying soil, and that Dactylella ellipsosporum can be controlled by Dactylella ellipsosporum.
There are two research reports in which control was achieved using P. lipsospora), but in most other research reports, no control effect was observed.

The present inventor believes that one of the reasons why no control effect has been observed is that in conventional research, bacteria that grow well on the culture medium are arbitrarily selected and used, ignoring the predation properties of nematode-predating bacteria. I thought that this was because I was hoping for a pest control effect.

Therefore, the present inventor has conducted research with a research policy that focuses on the predation characteristics of nematode-predating fungi, and has discovered that they have extremely excellent properties as natural enemies of the nematode nematode, and have hitherto unknown physiological effects. They discovered a new species of nematode-predating fungus with ecological specificity.

In other words, as part of our research, the present inventor is working to find the resistance factors that are suppressing the invasion and reproduction of the pine tree nematode in the bodies of resistant pines that have not suffered any blight damage. During this process, a nematode-predatory fungus discovered in the sap of healthy Japanese red pines has the property of selectively and rapidly hunting down pine nematodes.
It has the property of allowing hyphae to invade the tree and continue to live, and furthermore,
We confirmed that it is a new species that has the property of catching and killing pine nematodes in dead water and has various mycological properties that were unknown until now, and we established methods for cultivating it, methods for its application, etc., and developed the present invention. The method has been completed.

The method of the present invention is a method for preventing the blight of powder, which is characterized by applying Arthrobotrys sp. TKBC1015, which is a nematode-predating fungus belonging to the genus Arthrobotrys, to the powder. This is an epoch-making method in that it has a preventive effect against pine nematode disease and prevents blight, and when applied to dead trees, it has an exterminating effect on pine nematode and prevents blight.

Next, the configuration and effects of the method of the present invention will be explained in detail.

First, the outline and mycological properties of Arthrobotrys sp. TKBC1015 (Feikokenkyo No. 5648) used in the method of the present invention will be explained.

a. Growth status in each medium Malt extract agar medium: Growth is relatively slow at 25°C, fluffy and diffuse, and white to milky white.

The underside is pale yellowish white.

Potato glucose agar medium: Growth is almost the same as on malt extract agar medium, but the formation of aerial mycelia is the best.

Fluffy, diffuse, white to milky white.

The underside is pale yellowish white. Czapek agar medium: Poor growth and almost no formation of aerial mycelia.

b. Morphology, comparison, and identification Hyphae are irregularly branched, 2 to 3.5 microns wide, and colorless.

Conidiophores arise from aerial hyphae, have septate walls, are rarely branched or have pseudoaxial branches, and have a width of 3.0 to 3.0 mm.
5 microns and 170 microns in height, with conidial bulges of 3.5 to 3.8 microns wide in places in the middle or at the tips.

The conidia are white, symposio-shaped, and arise one by one from a small protrusion at the tip of the conidiophore, and as these small protrusions are produced one after another, the tip becomes enlarged as a result of the formation of several conidia.

Furthermore, the conidiophores elongate and form conidia in the same way, and the tips become enlarged, and several conidia gather in each enlarged part.

Conidia (spores) oblong, 2 cells, 14-17.5X
4-4.8 microns, colorless.

In the presence of pine needle nematodes, small stalks are produced from the sides of the germinated hyphae of basal hyphae and conidia, and the slimy substance formed at the tips selectively captures pine needle nematodes in a mochizao manner.

Based on the above-mentioned morphological properties, this bacterium has been identified in Illustrated Generals of Imperfect Fungy by Barnett Hunter (1972) and Hyphomycetes by Kendrick Carmichael, edited by Ainsworth Summerman, Fangyle A1 Academic. According to Press (1971), it is clear that it belongs to the genus Arthrobotrys in the subphylum Deuteromycota and the class Deuteromycetes.

The species closest to this fungus is Barron (BarronlG, L,
1977 The nematode destruction
ng fungi, Canadian Biol
logicalPublication, Guelph
, Ont, ), Sienck-Kendrick-Prama (5ohenck, Sl, Kendrick, W
, B1, & Pramer, D., 1977A n
ew nematode -trapping lyp
hamycete and a revision
of Dactylaria and Arthrob
otrys, Can, J, Bot, 55:9
77-985), there is Arthrobotrys superba, but the conidia of this bacterium are much smaller and do not match.

In addition, there is no species in the same genus that traps pine nematodes in the same manner as this fungus, and whose conidial traits match.

Therefore, it is certain that this bacterium is a new species included in the genus Arthrobotrys, and it has been named "Arthrobotrys sp. TKBC1015j."

C8 physiological properties Optimal growth conditions: pH 7.2, temperature 25°C.

Growth range: pH 2.0-9.0, temperature 18°-30
℃.

d. Other characteristics Adding about 0.5% yeast extract to the medium promotes growth.

After the growth of vegetative mycelium, lowering the culture temperature from 25°C to 20°C promotes spore production.

This bacterium was isolated from the sap of a healthy red pine tree and purified using conventional methods.

The pure isolated bacterium shows no physiological or morphological changes even in the 25th generation (after 1 year and 6 months) of subculturing with conidia, and is genetically stable.

The method of the present invention is carried out using Arthrobotrys sp.
It is simply called "T bacterium."

- is applied to powders to prevent them from withering.

Next, the means for cultivating T bacteria and the means for applying them will be explained.

Means for culturing T bacterium: The medium may be either a synthetic medium or a natural medium as long as it contains carbon sources, nitrogen sources, inorganic substances, and trace amounts of nutrients required by T bacterium.

Glucose, fructose, sucrose, etc. are used as carbon sources,
Ammonium salts, nitrates, etc. are used as nitrogen sources.
Growth is also good if less than 1% of yeast extract or Ebios (trade name) powder is added as a trace nutrient source.

The optimal culture temperature is 25°C, but it grows well within the range of 23°C to 26°C.

It grows well in a pH range of 55 to 7.5.

Either solid culture or liquid culture may be used.

In the case of solid culture, conidia are easily formed, and conidia are resistant to environmental changes, so they are advantageous for field application.

In the case of liquid culture, a large amount of bacterial cells can be obtained in a short period of time, but
Since the number of endospores is small, if it is necessary to obtain a large amount of spores, the cultured bacterial cells may be transferred to a solid medium using a liquid culture method and then cultured for several days.

In addition, the cultivation method suitable for short-term and large-scale cultivation of T bacteria is
This method uses white rice, rice bran, wheat bran, oil residue, soybean residue, monosaccharide millet residue containing okara, rice straw, rice hulls, etc. alone or in combination.
This method uses a medium containing 5 to 50% water, or a medium containing 10% or less sugars and 1% or less yeast extract. About 14 days after inoculation, the medium is filled with hyphae and aerial spores are formed, and when the culture is left at 20°C, the number of spores formed increases significantly.

Even if the T bacterium is cultured by any of the above-mentioned culturing methods, there is no physiological or morphological change in the spores or mycospores, and of course, there is no change in the ability to feed on pine nematodes.

Means of application of T bacteria: As a means of application, 1. Spray application, 11. Soil application, 1
11. Three means of tick-mediated application are suitable.

1. Spraying application In this case, it is preferable to use a spore suspension of T bacteria.

The spore suspension solution can be prepared by stirring the culture (medium) cultured by the various culturing methods described above using a stirrer, and then passing it through a cloth or the like to obtain a spore suspension solution.

Adjust the concentration of spores in the solution to an appropriate level (usually about 200,000 spores per 1 ml), and use it for foliar spraying.

A spreading agent may be added to the spore suspension solution if necessary.

Of course, when culturing is performed using an agar medium, such as P, D, or A medium, a spreading agent is not necessary since the spore suspension is an agar solution.

It is also possible to store the spores in the spore suspension solution by adsorbing them to a desiccant such as talc in a conventional manner, and dilute them with water before use.

The spore suspension is applied to the leaves, trunks, and roots of the target powder using a watering can, sprayer, sprinkler, etc.

Il. Soil application In this case, it is preferable to use a mass-cultured culture - the medium itself - and apply it to the soil near the roots of the target powder by spreading it underground to about 771 degrees. .

It is particularly preferable to apply koji as it is on which a large amount of T bacterium spores have been formed by the above-mentioned "koji culture method."

111, application using ticks as vectors.

Several types of mites have been found in the tubs, cavities, and adult bodies of the Matsuno-dachi longhorn beetle, and it is known that these mites are natural enemies that prey on nematodes, and a method for culturing them has already been established. A magazine called "Forest Epidemic Prevention J, February 1980 issue, P36-P39-0. This mite searches for the tubs, cavities, and adult bodies of the pine beetle in dead water, so it acts as a carrier for the T bacterium. Available.

According to the research of the present invention, each adult tick can carry about 150 spores of T bacteria.

The spores of the T bacterium used in this case can be prepared by culturing the spores on solid culture or on paper impregnated with a culture solution and collecting them by scraping them off the culture surface with a brush.

If the spores prepared as described above are placed in an incubator in which mites are being cultured and thoroughly stirred, the spores will be deposited on the mites.

If you release these mites into areas where target plugs grow,
The released mites carry their spores intensively to the capsule and body of the pine beetle in the tree.

The spore suspension, dried spores, culture medium, and collected spores prepared by the application methods in 1 to 111 above can be stored at room temperature, but if stored for a long period of time, it is necessary to store them in a cool, dark place at about 5°C. It is desirable to store it in

The means for applying the T bacterium is not limited to 1 to 111 above, and for example, the T bacterium is inoculated into a culture medium containing appropriate moisture, cultured, and the like. It can also be applied by wrapping it around the trunk of a plug tree, which is intended for this purpose.

Next, we will explain the safety of T bacteria. First, as mentioned above, the growth temperature range of T bacteria is 18° to 30° C.
Since it does not grow at 0°C and at human body temperature, it will not grow even if it enters the human body, and safety for humans can be guaranteed.

(The inventor has been handling T bacteria with bare hands for over a year, and has not noticed any abnormalities.

) Furthermore, the present inventor has confirmed the safety to animals and plants through the following tests.

Safety test for O. silkworm; Spores on the bacterial layer of T bacterium cultured on agar medium (P, D, A medium with 1/10 concentration) were treated with surfactant (Aeroso
l OT: Trade name) 0.1% agar aqueous solution containing 0.001% was suspended and collected, and approximately 25 spores were collected per ml of agar aqueous solution.
A spore suspension prepared to contain 0.00 spores was sprayed onto collected leaves using a sprayer, and the collected leaves were fed to 3 of the 4 silkworm larvae.

All three of these silkworms metamorphosed into pupae, became adults, and laid eggs, but during this process, there were no abnormalities compared to control silkworms that were fed normal leaf collection without spraying the spore suspension. This was not observed at all, confirming that T bacteria did not affect the growth of silkworms.

Safety test for O radishes and rapeseed; Prepare 4 pots filled with field soil, sow 10'0 radish seeds in each of the 2 pots, - pots in the test area, and the other pot. The two pots were used as a control, and each of the two pots was sown with 100 rapeseed seeds.
One pot was used as a test plot, and the other pot was used as a control plot.

In each test plot, the same T bacterium spore suspension used in the silkworm test was sprayed every day from the day the seeds were sown using a sprayer, and the germination rate was measured on the 10th day.

In addition, water was sprinkled on the control pots every day from the day the seeds were sown, and the germination rate was also measured on the 10th day.

The germination rate was 100% in both the test plot and the control plot, confirming that T bacteria had no effect on the germination rate.

Next, some of the many experimental examples conducted by the present inventors will be extracted to explain the effect of T bacterium on preventing plugs from withering.

Experimental example 1 [Application to healthy trees] Healthy red pine 3-year-old seedlings (height: average approximately 30C7rL,
The seedlings (weighing on average about 351) were transplanted into plastic No. 6 pots containing 2501 field soil in early April, and the seedlings that had sprouted two months later were used in this experiment.

There were 30 pots used in the first experiment, and one seedling was planted in each pot.

Of the 30 pots, 10 were treated, 10 were control, and the rest were comparison.

Agar medium (P, D,
The Botrytis nematode grown on Botrytis fungi on A medium) and washed with sterile water was inoculated by the following procedure.

Split the branch tip of each seedling with scissors and fill the split with absorbent cotton to serve as the inoculation point.

Three inoculation points are formed for each seedling, and about 3,300 pine white nematodes are inoculated in a liquid form at each spot, so that each seedling is inoculated with a total of about 10,000 pine white nematodes.

The vaccination was carried out in mid-July.

In the treatment area, use a sprayer to spray the same T fungus spore suspension solution used in the silkworm test at a rate of 2ml/1l per seedling on the entire exposed part of the seedling and on the soil surface of the pot. do.

The above-mentioned spraying was carried out once a week for two months from mid-June, one month before inoculation with pine nematode, to mid-August, one month after inoculation.

The control plot was sprinkled with tap water as appropriate.

In the comparison area, tap water containing 1,100 ppm of sodium nitrate and 10 ppm of dipotassium phosphate was administered at a rate of 100 ppm once from mid-June, one month before inoculation with pine nematode, to mid-August, one month after inoculation. The punishment was twice a week for two months.

The condition of each seedling 2 months and 6 months after inoculation with pine nematode was as shown in Table 1.

In the above table, the state in which the seedlings wither and die is displayed in five stages, and the seedlings whose whole body is red and withered are marked as dead,
Seedlings that are mostly red and withered but with some green leaves remaining are called "almost dead," seedlings that have about the same ratio of green and red leaves are called "half dead," and seedlings that have only a few dead red leaves are called "almost dead." Seedlings that were partially dead and remained healthy were healed.

From the above experimental results, it was confirmed that the spraying of T bacterium spores is effective in preventing the death of healthy trees, and is significantly superior to the effect of preventing the death of healthy trees when compared with the use of chemicals.

Experimental Example 2 [Application to Dead Trees] Dead trees of Akamatsu en 146 on the campus of the University of Tsukuba in 1972 were used in this first experiment.

Two trunks of this dead tree were cut into lengths of 1.3 m, one for treatment and one for control.

Spores on a strain of T bacterium cultured for one month on agar medium (1/10 concentration P, D, A medium) were treated with a surfactant (Twe
en-80: trade name) was suspended in a 3% saline solution containing 0.001% to prepare a saline solution containing about 200,000 T bacterium spores per ml, and 5ml of it was collected using a sprayer for treatment. Sprayed all over the log.

On the other hand, control logs contained 3 containing 0.001% surfactant (Tween-80: trade name).
% saline solution was sprayed all over the log using a sprayer.

The treatment and control logs were sprayed on May 31st.

After spraying, we collected the pine beetle that emerged and escaped from both logs, separated the pine beetle using Behrman's method, and measured the number of nematodes that migrated.

From June 4th to June 19th, 6
Three heads emerged from the control log between June 10th and June 18th.

The numbers of pine nematodes held by these pine beetles are shown in Table 2.

As shown in Table 2, pine beetle nematodes have no longer migrated to the pine beetle that emerged and escaped 19 days after the T fungus spores were sprayed.

This is presumed to be the result of the pine nematode inside the log being captured and killed by the T bacterium.

In addition, when the log for processing was cut and observed under a microscope, it was observed that mycelium of T bacterium was growing inside the tissue.

From the above experimental results, it is possible to exterminate the pine nematode by spraying T fungus spores on dead trees, and also to interrupt the life cycle of the pine nematode, which uses the pine beetle as a vector, in the pine wilt mechanism described above. was confirmed.

Experimental Example 3 [Application on a weakened tree] This experiment was conducted on a weakened red pine tree approximately 25 years old on the campus of the University of Tsukuba.

According to the sap determination method, the above-mentioned weakened tree is a weakened tree that does not exude sap at all.Appearance-wise, the leaves are the same green color as a normal healthy red pine, but some branches are wilted. is in a state of

We carried out Behrman's Civil Law on the above-mentioned weakened trees, but no pine nematode was detected.

However, judging from the environment, this weakened tree was presumed to be in the early stages of being infected by pine nematode, and was selected as the subject of this experiment.

A platinum loopful of T bacterium spores was inoculated onto a plain agar medium with a diameter of 9 CrrL and a thickness of 5 mm, and cultured at 24° C. for 3 days.

This agar medium was directly attached to the cambium of the chest height region of the above-mentioned weakened tree (the outermost part of the material from which the bark was peeled off) and fixed with tape.

After leaving the tree in this state for 24 hours, the medium was removed from the surface of the tree and the surface of the medium was observed under a microscope, and it was found that three pine nematodes were captured by T bacteria on the medium.

From the above experimental results, it was confirmed that the T bacterium has the ability to attract and kill pine nematodes from weakened trees in the early stages of infection when the pine nematode density is low.

As is clear from the above experimental results, the T bacterium used in the method of the present invention can survive inside trees, whether they are healthy trees, dead trees, or weakened trees, and has the ability to capture and kill pine tree nematodes inside trees. It is effective in preventing powders from withering.

Furthermore, as seen in Experimental Example 3, T bacterium has the ability to attract and kill pine nematodes in weakened trees in the early stages of infection, and uses this ability to attract and kill pine nematodes in weakened trees in the early stages of infection. It is also possible to perform early detection of

That is, a patch-shaped medium containing T fungus spores is prepared, and this is pasted on each tree body in a pine forest, and after a predetermined period of time (for example, 24 hours), it is removed to determine whether or not the T fungus has killed the pine tree nematode. By examining a tree under the microscope, it is possible to determine whether or not it is infected at an early stage, even if the tree is in the early stages of infection, which cannot be detected under the Behrmann Civil Code.

According to the method of the present invention as explained above, the fundamental extermination of the pine nematode, which was considered impossible in the past, can be easily carried out, and the blight of powder can be effectively prevented.

Example 1 "Koji culture method" using soybean waste - 4 kg of soybean waste
, water 1. G, 24°C for 115 days to culture T bacterium - Kaikoken No. 5648 - in 5 kg of medium containing approximately 1,000,000 T bacterium spores per 1 g (Durham) medium.
was prepared.

A healthy red pine that is about 25 years old and uninfected by T bacteria (location:
5 kg of the above T bacterium culture (medium) was applied to an isolated tree with a diameter of 25 cm at breast height and a height of about 7 m at Shishido International Golf Club, 1340 Minamikoizumi, Tomobe-machi, Nishiibaraki-gun, Ibaraki Prefecture.

The application was carried out by digging into the ground in the area covered by the above-mentioned isolated tree technique to a depth of about 5 tsp.

The date of application was March 25, 1982.

As of June 1, 1980, the above-mentioned isolated tree is growing in a healthy condition without any obstacles.

The following inspections were conducted on the above-mentioned isolated trees on June 1, 1980 and April 20, 1981.

That is, the roots of the above-mentioned isolated tree, the bark at the ground edge, the sap collected from the chest height, the leaves, and the soil at the base of the above-mentioned isolated tree are taken as samples, and each sample is placed in the center of the agar medium. Approximately 100 pine nematodes were inoculated into the surrounding area, and after culturing at 24°C for one week, each sample was examined for the presence or absence of T bacterium.

As a result, T bacteria appeared with a probability of 10% from the roots, 60% from the sap, and 10% from the soil, but no T bacteria appeared from the bark or leaves.

Furthermore, according to the sample results on April 20, 1981, the detection rate of bacteria was 10% in roots, 20% in sap, and 0% in other areas.

These results indicate that by applying the T bacterium culture to the ground as described above, the T bacterium invades the body of the isolated tree, survives within the tree, and maintains the ability to kill pine tree nematodes. can be confirmed.

Example 2 Medium 1' prepared by the same method as Example 1? 20 kg of medium containing about 1,000,000 T fungus spores each
and 1 ml of spore suspension solution prepared by the following procedure.
A spore suspension solution 15007 containing about 2500 T bacterium spores per sample was used.

For 100 red pine trees in a 25-year-old red pine forest (location: Shimohara Kasumigaura International Golf Course, Yatabe-cho, Tsukuba-gun, Ibaraki Prefecture), the above medium was applied in the same manner as in Example 1, at a rate of 0.2 kg per tree. The above culture solution was applied to leaves and tree trunks using a sprayer in an amount of 151 per plant.

The date of application was May 13, 1981.

Of the 100 red pines, 5 were inoculated with about 30,000 pine nematodes per tree on June 3, 3 weeks after the treatment.

As of June 1, 1981, the above 100 red pines are:
All but one of the plants that were inoculated with Atsunosai nematode are growing without withering.

The spore suspension was prepared as follows.

Glucose 101, yeast extract 1.5g, MgSO4.
71 (2Q0.5g, N'lH4NO30.5g, K
2HPO40.5P, NaH2PO40.4? A culture solution (pH 6,8) with a composition adjusted to 11 with tap water was inoculated with T bacterium - Microtechnical Research Institute No. 5648, and cultured at 24°C for 10 days, and the culture solution was stirred well using a stirrer. After that, I passed the river,
A spore suspension solution 41 containing 1,000,000 T bacterium spores per ml is obtained.

Water was added to this hanging solution to adjust it to a solution containing 2,500 spores per 11, and CMC glue was added as a spreading agent to 0 per 11.
．． 31 and use it.

Claims (1)

[Claims] 1 Arthrobotrys sp. TKBC1015 (A
rthrobotrys sp, TKBC1015)
A method for preventing blight of powders, characterized by applying -Feikokenkyori No. 5648- to powders.