The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 11,493 M/T in 2014. Several fungus are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Cladobotryum mycophilum is the causal agent of cobweb disease of commercial mushrooms. Early symptoms were noticed as round, fleshy, yellowish brown lesions on mushroom caps. Late symptoms progressed when the parasitic fungus formed white cobweb circular colonies on dead or damaged pinheads, spread on the surface of the casing, and covered entirely fruiting bodies. A Gram-positive bacterium was isolated from mushroom media that markedly showed the antagonistic activity against Cladobotryum mycophilum, the most destructive pathogen of cultivated mushrooms. The HC57 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus subtilis by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for Cladobotryum mycophilum cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Cladobotryum mycophilum. Control efficacy of browning disease of strain HC57 treatment was 71% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 1.5% Xylose, 2% Soytone, 1% NH4H2PO4, 7 mmol CaCl2, and 0.5% Histidine at pH 6.0 at 25℃. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by Cladobotryum mycophilum.

Mushroom is cultivated as one of the major economical crops in many areas in Korea. The total production has steadily increased from approximately 198,563 M/T in 2009 to 208,941 M/T in 2012. Several fungus are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Trichoderma harzianum is the causal agent of brown blotch disease of commercial mushrooms. Colonization of mushroom caps by the bacterium results in development of brown or cream lesions on pileus and stipe. These lesions are slightly concave spots and can be round or spreading. Antagonists against Trichoderma harzianum, CAM33 were selected and their control efficacy of green mould disease was investigated in this study. The CAM33 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus methyrotrophicus. by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for Trichoderma harzianum cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Trichoderma harzianum. Control efficacy of browning disease of strain CAM33 treatment was 77% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 3.0% Saccharose, 1.5% Soytone, 1.0% (NH4)2HPO4, 10 mmol MgSO4, and 2.0% Glutamic acid at pH 6.0 at 25°C. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by Trichoderma harzianum.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 10,996 M/T in 2012. Several fungus are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Cladobotryum mycophilum is the causal agent of cobweb disease of commercial mushrooms. Early symptoms were noticed as round, fleshy, yellowish brown lesions on mushroom caps. Late symptoms progressed when the parasitic fungus formed white cobweb circular colonies on dead or damaged pinheads, spread on the surface of the casing, and covered entirely fruiting bodies. A Gram-positive bacterium was isolated from mushroom media that markedly showed the antagonistic activity against Cladobotryum mycophilum, the most destructive pathogen of cultivated mushrooms. The HC7 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus altitudinis. by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for Cladobotryum mycophilum cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Cladobotryum mycophilum. Control efficacy of browning disease of strain HC7 treatment was 78% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 3% Soluble startch, 10% Soytone, 1% (NH4)2HPO4, 1 mmol KCl, and 0.5% L-asparagin at pH 6.0 at 30°C. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by Cladobotryum mycophilum.

Mealworm, Tenebrio molitor L. (Coleoptera: Tenebrionidae) has high and safe protein contents, which enables it to be animal feed. However, occurrence of entomopathogenic fungi in mealworms is one of the limitations for mass production. In this work, we investigated relationships between abiotic conditions and occurrence of fungal pathogens and established an effective control method using fungicides. In virulence assay, third instar mealworm larvae were sprayed by six entomopathogenic Beauveria bassiana isolates and kept under high relative humidity; B. bassiana ERL1575 isolate had highest virulence. Under normal humidity, ERL1575 conidia showed different virulence between spray (~0% virulence) and digestion (~80% virulence) method. Furthermore, mealworms, which digested conidia, were exposed to various temperature (20-35°C) and humidity (1-3 ml distilled water spray/35 mm diam. dish) conditions for 5 days. All the treatments showed ~90% virulence except 35°C incubations (~20% virulence), but irrespective to the humidity conditions. Forty chemical fungicides were assayed against conidial germination and hyphal growth of ERL1575. Fluazinam and mancozeb showed strong inhibition of conidial germination at standard application dose (SD), 1/2 SD and 1/5 SD; besides, fluazinam showed strong inhibition of hyphal growth. When fluazinam and mancozeb were applied to the fungal conidia-inoculated wheat bran, most of mealworms were alive after 3 days post application. However, high mortality rate (~100%) were observed in the conidia-inoculated wheat bran without any fungicides. In conclusion, this work suggests that B. bassiana isolates could be pathogens at <30°C when they were digested by mealworms, and fluazinam and mancozeb would be used as effective control agents against the pathogen.

고추 역병은 그 병원균의 토양전염성 때문에 약제에 의한 방제효과가 낮아 저항성 품종의 개발이 기대되어 왔다. 고추 역병에는 CM334, AC2258, PI201234 등 다수의 저항성 유전자원이 보고되었으며, 이들의 저항성의 유전에 관한 연구로 진행되었다. 그러나 저항성의 유전양식은 실험에 사용한 재료와 연구자에 따라 1개, 2개, 혹은 3개 이상의 유전자, 다수의 유전자에 의한 양적 유전 등 다양하였다. 최근에는 분자적 방법으로 양적형질유전자좌(QTL)를 구명하는 연구가 보고되고 있으며, 분자표지를 이용한 선발 기술도 이미 육종 현장에서 활용되고 있다. 최근 저항성 품종이 다수 출시됨에 따라 새로운 병원형(pathotype), 즉, 레이스(race)의 출현에 관심이 높아지면서 이에 대한 연구가 보고되고 있다. 모두 품종과 병원균주간에 특이적 변이가 있으며, 이를 토대로 몇 개의 병원형(race)으로 분류할 수 있었다. 그러나 판별품종이 통일되지 않았고 시험에 사용한 품종들의 저항성 유전자의 조성도 달라 세계적으로 통일된 레이스분류체계는 아직 없는 실정이다. 이러한 배경에서 보다 안정된 저항성 품종의 육성을 위해서는 한 가지 저항성 재료보다는 다수의 저항성 유전자원에서 저항성을 도입하고, 육성과정에 육성품종의 보급 대상지역의 여러 균주를 사용하여 선발하는 것이 필요할 것이다.

The button mushroom, Agaricus bisporus, is one of the major economical crops cultivated in Korea. This mushroom showed the 5th production to 10,996 M/T in 2012. Several bacteria are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Pseudomonas agarici is the causal agent of browning disease of commercial mushrooms. Colonization of mushroom caps by the bacterium results in development of browning lesions on pileus. These lesions are superficial brown spots and can be round or spreading. But P. agarici never caused sunken lesions and rotting of the mushroom tissues. A Gram-positive bacterium was isolated from mushroom media that markedly showed the antagonistic activity against Pseudomonas agarici, the most destructive pathogen of cultivated mushrooms. The HC42 strain was selected as antagonistic bacterium by inhibition zone method and it was identified as Bacillus safensis. by the cultural, morphological and physiological characteristics, and analysis of the 16S rDNA.. The isolated bacterium is saprophytic but not parasitic nor pathogenic to cultivation mushroom. The isolated bacterium for P. agarici cell, was sufficient for inhibition in vitro. Inoculation of the isolated bacterium prevents the development of bacterial disease in Agaricus bisporus. Control efficacy of browning disease of strain HC42 treatment was 66% on Agaricus bisporus. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 1.5% D-galactose, 1.5% yest extract, 1% NH4Cl, 1.5% KCl, and 1.0% L-asparagin at pH 6.0 at 25℃. The suppressive bacterium may be useful in future for the development of biocontrol system and the construction of genetically modified edible fungi resistant to the disease caused by P. agarici.

Mushroom is cultivated as one of the major economical crops in many areas in Korea. The total production has steadily increased from approximately 186,400 M/T in 2007 to 190,111 M/T in 2011. The button mushroom, Agaricus bisporus, showed the 5th production to 13,052 M/T in 2011. Several bacteria are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Pseudomonas agarici is the causal agent of browning disease of commercial mushrooms. Colonization of mushroom caps by the bacterium results in development of browning lesions on pileus. These lesions are superficial brown spots and can be round or spreading. But P. agarici never caused sunken lesions and rotting of the mushroom tissues. Antagonists against P. tolaasii, HC42 were selected and their control efficacy of browning disease was investigated in this study. Antagonists against P. agarici, HC42 were selected and their control efficacy of browning disease was investigated in this study. After proceeding antagonistic test, HC42 was selected as a strong antagonist against P. agarici and the HC42 strain was identified as P. safensis with the cultural, physiological and biochemical properties and analysis of the 16S rRNA. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 1.5% D-galactose, 1.5% yest extract, 1% NH4Cl, 1.5% KCl, and 1.0% L-asparagin at pH 6.0 at 25℃. Control efficacy of browning disease by HC42 treatment was 66% on Agaricus bisporus..

Mushroom is cultivated as one of the major economical crops in many areas in Korea. The total production has steadily increased from approximately 186,400 M/T in 2007 to 190,111 M/T in 2011. The button mushroom, Agaricus bisporus, showed the 5th production to 13,052 M/T in 2011. Several bacteria are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Pseudomonas agarici is the causal agent of browning disease of commercial mushrooms. In this study, antagonistic bacteria to P. agarici were selected in vitro tests using confrontation bioassay and paper disk diffusion assay. The most active bacteria, HC12 were selected and their control efficacy of brown blotch disease was investigated in this study. After proceeding antagonistic test, HC12 was selected as a strong antagonist against P. agarici and the HC12 strain was identified as Alcaligenes sp. with the cultural, physiological and biochemical properties and analysis of the 16S rRNA. The optimal culture medium for the antagonistic bacteria growth was determined as follows: 0.5% dextrin, 1.5% yest extract, 1% NaNO3, 0.5% KH2PO4, and 1.5% L-asparagin at pH 9.0 at 30℃. Control efficacy of browning disease by HC12 treatment was 63% on Agaricus bisporus..

Mass mortality of oak trees has been reported in Korea since 2004. It seemed to be occurred by a possible pathogenic fungus Raffaelea quercus-mongolicae and spread by an ambrosia beetle, Platypus koryoensis (Coleoptera: Platypodidae) which was a vector of that fungus. P. koryoensis attacked healthy or stressed living Quercus mongolica. The numbers of damaged and/or died Q. mongolica by P. koryoensis has increased every year since its first report. The damages caused by Platypus spp. were reported world widely. Pinehole borers P. caviceps, P. apicalis and P. gracilis damaged and killed living southern beech (Nothofagus spp.) and/or Kamawi (Weinmannia racemosa) in New Zealand. The relationship between pinhole bores, the fungal pathogen Sporothrix and their host beech is well established. P. subgranosus was responsible to death of myrtle beech (N. cunninghamii) in Tasmania, Australia. In southern Europe, P. cylindrus attacked cork oak (Quercus suber), and Phytophthora cinnamoni was known as a pathogen. P. mutatus was a primary pest to poplars, especially Populus deltoides in South America. Alfaro et al. warned on the threat of P. mutatus to world poplar resources since its introduction to Italy on 2002. P. quercivorus, a vector of pathogenic fungus Raffealea quercivora was responsible for Japanese Oak wilt disease on Q. crispula and Q. serrata. In this study, we will introduce the current status of oak wilt disease in Korea and integrated control method of this disease

Mushroom is cultivated as one of the major economical crops in many areas in Korea. The total production has steadily increased from approximately 186,400 M/T in 2007 to 198,563 M/T in 2009. Several bacteria are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Pseudomonas tolaasii is the causal agent of brown blotch disease of commercial mushrooms. Colonization of mushroom caps by the bacterium results in development of brown or cream lesions on pileus and stipe. These lesions are slightly concave spots and can be round or spreading. Antagonists against P. tolaasii, HC1 were selected and their control efficacy of brown blotch disease was investigated in this study. The HC1 isolate was selected as an inhibitor of tolaasin activity by bioassay on potato and it was identified as Pseudomonas sp. by the cultural, physiological and biochemical properties and analysis of the 16S rRNA. Control efficacy of brown blotch disease by HC1 treatment was 69% on Agaricus bisporus, 68% on Flammulina velutipes and 55% on pleurotus ostreatus respectively.

The total production has steadily increased from approximately 186,400 M/T in 2007 to 198,563 M/T in 2009. Winter mushroom, Flammulina velutipes, with 61,057913 M/T in 2009, showed the highest production. Several bacteria are known as the causal agents of diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). Pseudomonas tolaasii is the causal agent of brown blotch disease of commercial mushrooms. Antagonists against P. tolaasii, HC5 were selected and their control efficacy of brown blotch disease was investigated in this study. After proceeding antagonistic test, HC5 was selected as a strong antagonist against P. tolaasii and the HC5 strain was identified as P. azotoformans with the cultural, physiological and biochemical properties and analysis of the 16S rRNA. Control efficacy of brown blotch disease by HC5 treatment was 73% on Agaricus bisporus, 78% on Flammulina velutipes and 71% on pleurotus ostreatus respectively.

Pine wilt disease, caused by pine wood nematode, Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle, has become the most serious threat to pine trees in Korea since first reported in 1988. Pine wood nematode is transferred to uninfested trees by Monochamus alternatus (Coleoptera: Cerambycidae) (vector). A typical controlling method against vectors in Korea is fumigation of the dead trees infested by vector M.alternatus and B.xylophilus using metam-sodium SL(25%). However, this method is harmful to the environment because of the chemical contamination and vinyl waste in the mountain afterward. Portable Bark Remover (PBR) was contrived to reduce these environmental problems. The vectors oviposite under the bark of the freshly dead trees only. Debark infested trees prohibited the vectors from oviposite and finally, vectors can not complete their life cycle. The PBR is a newly designed as a debarking device that is equipped on top of the chain saw engine allowing ease and rapid debarking of the infested trees at the controlling field. Debarking of freshly-dead trees can eliminate egg-laying and hatch places where between the wood and bark from vectors. The new method for vector controlling using the debarking device is expected to lower-price and high-efficiency comparing with other conventional methods such as “crushing”, “burning” and “fumigation”.

This study was carried out investigate to availability of commercial microbial pesticide and antibacterial activity of isolates isolated from different mushroom media. Ten commercial microbial pesticide and EM liquid were collected from different company. EM of these one has been used for control of mushroom disease and growth promotion at the mushroom farms. The density of bacteria and yeast in EM cultural liquid was higher than those of the crude liquid. The pH values of EM showed the low acid levels(pH 3.5~3.9) by organic acid secreted from microorganism. The kinds of organic acid was acetic acid, lactic acid and succinic acid. The dominant bacteria isolated from EM liquid was Lactobactillus sp.(21 strains), Acetobacter sp.(9 strains), PaeniBacillus sp.(9 strains) and others(12 strains). The organic acid bacteria isolated from fermentation foods( was 92 strains and the dominant genus was Weissella sp.(41 strains), Leuconostoc sp.(21 strains), Enterococcus sp.(9 strains), Lactococcus sp.(9 strains) and others(12 strains). And we isolated 2,500 bacteria from oyster mushroom and button mushroom cultural media for selection of antagonistic bacteria. Thirty five strains of these isolates showed very strong antagonistic activity. These strains were identified Bacillus amyloliquefaciens, Bacillus subtilis, Brevibacterium halotolerans, Pseudomonas libanensis, Stenotrophomonas maltophilia and Alcaligenes faecalis by 16S rDNA analysis.

Several bacteria have been known as the causal agents of certain diseases of the cultivated button mushroom (Agaricus bisporus) and oyster mushroom (Pleurotus ostreatus). It is well known as bacterial diseases of the cultivated mushroom such as brown blotch, mummy disease, bacterial pit, bacterial rot and weeping disease, ginger blotch, and drippy gill. Brown blotch is the most critical cause of crop loss in the commercial mushroom industry. The classical bacterial blotch disease of mushrooms is caused by a fluorescent pseudomonad, Pseudomonas tolaasii. Affected mushrooms show lesions which become dark chocolate-brown, are wet, and deeply pit the caps and stalks. Although Pseudomonas tolaasii has been known as the casual agent of bacterial blotch, much controversy exists regarding the identification of this bacterium and whether blotch may be caused by more than one organism. This study was carried out to investigate characterization and biological control of Pseudomonas tolaasi and other possible browning pathogens isolated from cultivated mushrooms. One hundred seventy four bacteria were isolated from the cultivated mushroom and collected from main producing districts throughout the country. The isolates were classified into Pseudomonas tolaasii(20 strains), Pseudomonas gingeri(1 strains), Pseudomonas agarici(4 strains), Pseudomonas putida(11 strains), Pseudomonas sp.(46 strains), Ewingella americana(14 strains), Stenotrophomonas sp.(4 strains), and others(74 strains) on the basis of 16 rDNA analysis. The most dominants of these species were Pseudomonas tolaasii and Ewingella americana. Pseudomonad isolates were mainly divided into two groups in white line test and a sharply defined white line of precipitate forms in Pseudomonas agar F(Difco) between the opaque white colonies of P. tolaasii and translucent colonies of certain unidentified pseudomonads. The white line test was positive when 20 isolates of P. tolaasi from different countries were examined, whereas 62 isolates of pseudomonads did not give the white line reaction with a reacting translucent colony Pseudomonas. All the isolates tested for white line forming bacteria including P. tolaasi were highly pathogenic to mushroom tissue. Although browning of mushrooms in host tests does not perfectly help in the identification of P. tolaasi, a conspicuous pitting produced at the cut surface of mushroom tissue is as specific as the white line test in detecting P. tolaasii in suspension in distilled water. URP2F primers of 20-mer were used to assess the genetic diversity of white line forming bacteria. The phylogenetic tree was constructed by using the neighbor-joining method. In the analysis of RAPD pattern, all isolates of white line precipitate have some of the different genetic traits as collected districts. Phylogenetic analysis of 16S rDNA revealed that twenty isolates including white line forming bacteria were closely related to P. tolaasii and showed high similarity. To biological control on bacterial browning disease of cultivated mushrooms, six hundreds plant extracts (332 EtOH extracts, 268 water extracts) was used for control of mushroom disease. Thirty plant extracts in bacterial disease(Pseudomonas tolaasii, P. agarici, B. gladioli, E. americana) and thirty three in fungus disease(T. harzianum, C. mycophilum, V. fungicola) showed strong anti-microbes activity. They showed stronger anti-microbes activity at ethanol extracts than water extracts. MIC of extract BCW128 on Pseudomonas tolaasii was 700ppm and HDE17 was 330ppm. MIC of extract YCE107 on P. agarici was 330ppm, JGE96 was 330ppm and BCW128 was 700ppm. The bacteria inhibit tolaasin secreted by Pseudomonas tolaasii was selected three genus(Bacillus sp. etc). Now we are carrying out more research on these bacteria.

Pine wilt disease (PWD) represents a major threat to forest ecosystems worldwide. Although PWD is now better understood, effective control measures for this disease have still not been devised. Here, we report several years of field studies on preventative silvicultural control of PWD. Silvicultural control through preventative clear-cutting and the manual removal of logs was implemented between 2005 and 2009 in 16 Korean districts that had newly PWD-infected stands. Preventative clear-cutting of neighbouring asymptomatic pine trees (within a 10 to 50-m radius ofwilt trees) and the removal of felled logs or branches suppressed spread of PWD. Occurrences of PWD wilt pines in districts (city or county) subjected to this silvicultural control method were significantly reduced compared with those in districts using conventional controls (physical or chemical treatment of wilt pine trees). Through silvicultural control, PWD was successfully suppressed in 11 of 16 districts investigated. In contrast, successful control was achieved in only 1 among 18 districts subjected to conventional control. Our results will be of considerable interest to those engaged in the very difficult battle against the global spread of PWD.

It is well known that the root-knot nematodes, Meloidogyne spp., incite and aggravate the diseases caused by fungal and bacterial pathogens. The synergistic effects of the inoculation of Meloidogyne incognita combined with Fusarium oxysporum f. sp. lycopersici showed the greatly increased wilt symptoms developed on tomato plants compared to the inoculation of either of the two pests alone. For the biological control of the complex disease, a variety of bacterial isolates were tested for antagonistic effects to select ones that had both nematicidal and antifungal activities. Among forty plant growth-promoting rhizobacteria (PGPR) tested, Paenibacillus polymyxa G508 and G462 and P. lentimorbus G158 showed strong antifungal and nematicidal activities against F. oxysporum f. sp. lycopersici and second-stage juveniles (J2) of M. incognita, respectively, and also inhibited egg hatch of the nematode. The addition of Paenibacillus strains into potted soil suppressed the Fusarium-wilt severity and root galling on tomato and increased plant growths. P. lentimorbus G158 were abundantly proliferated on tomato seeds and hypocotyls more than P. polymyxa G 462 and had no phytotoxic effect on tomato plant. Under the greenhouse conditions, seed treatment of P. lentimorbus G158 reduced wilt severity caused by Fusarium wilt-root knot disease complex and root gall formation and increased tomato growth compared to the untreated control. Root-galls caused by both pathogens treated with bacterial culture had fewer and smaller giant-cells than untreated control, and scanning electron microscopy revealed alteration and distortion of hyphal cell wall of F. oxysporum and lysis of M. incognita egg shell by the bacterial treatment. All of these results suggest the Paenibacillus strains, especially G158 may have a high potential developed as biological control agents for the complex disease.

The most effective methods against pine wilt disease (Bursaphelenchus xylophilus, PWD), black pine bast scale (Matsucoccus thunbergianae), pine needle gall midge (Thecodiplosis japonensis) is the trunk injection of insecticides. The period of trunk injection that was considered with ecology and physiology of pests such as pine wilt disease, black pine bast scale and pine needle gall midge, was applied from Dec. to next Feb. and June, Dec. respectively. And there are differences in quantities of being injected chemicals to pine trees by the period of trunk injection. Thus, we investigated to prevent the effect of insecticides, to estimate the effect under the trunk injection of insecticides for pine trees against pests by the injecting period and time, boring site, opening and shutting of injection site. On Pinuns thunbergii, it was examined to figure out the effect of insecticides by the injecting period, twice a month on a sunny day around 10 AM from January to September. Injecting of insecticides was tested at a dose of 5ml per cm dbh of a pine tree after boring with hand drill at 50 cm high from the ground. As a result, abmectin 1.8% EC and phospamidon 50% SL were injected over 90% of mortalities from the end of January to the middle of March but as time goes by, injecting insecticides tended to decrease. To compare the volume of injecting insecticides in a day, between 10 AM and 2 PM on February 23rd, abamectin 1.8% EC was injected at a dose of 5ml per pine trees with micro pipette after boring with a drill machine at 50 cm high from the ground. As a result, injecting insecticide of a pine which was injected around 10 AM was completely injected and about 90% of the insecticide was injected in case of a pine that was injected around 2 PM. Trunk injection of insecticides was examined to figure out differences of the volume of injected insecticides before and after rain. There was no difference not only the rainfall but also the bored direction.