The entomopathogenic bacteria Xenorhabdus/Photorhabdus inhibit insect immune responses by inhibiting eicosanoid biosynthesis. Especially, the bacterial secondary metabolites inhibit PLA2 that release eicosanoid biosynthesis precursors. Some organic extracts of the bacteria-cultured broth possessed PLA2 inhibitory activity. This study used butanol to extract the metabolites and showed that the extract possessed potent inhibitory activity against insect immunity. The inhibition was reversed by the addition of eicosanoid biosynthesis precursor, suggesting PLA2 inhibition. Furthermore, sub-fractionation of the butanol extract separated the secondary metabolites and the fractions exhibited differential PLA2 inhibition. The active fractions appeared to contain novel compounds that are not known in PLA2 inhibition of these bacteria.

뿌리혹선충(Meloidogyne spp.)은 전세계 농업지대에 널리 분포하며 경제적 피해를 끼치는 선충 집단이다. 약 60여종의 뿌리혹선충 중에서 농업상 중요한 종은 M. arenaria, M. hapla, M. incognita와 M. javanica 등 4종이다. 식물의 뿌리에 침입한 유충은 생리활성물질을 방출하여 거대세포를 형성하며, 기주의 양분 및 수분 흡수기능을 저하시킨다. 우리나라에서는 땅콩, 들깨, 딸기, 토마토, 작약 등의 채소작물, 약용작물 및 기타 주요 경제적 작물들이 피해를 받고 있다. 시설하우스에서 재배되는 채소 및 화훼작물 또한 그 피해가 심하지만, 연용 시 높은 독성 및 토양 잔류성, 지하수 오염 등의 문제를 가지는 기존의 살선충제 외에 적절한 방제법이 없는 실정이다. 따라서 본 연구는 환경에 안전하면서도 효과적인 뿌리혹선충의 방제법 개발에 목적을 두고, 토양으로부터 Photorhabdus sp.를 분리‧이용하여 식물기생선충에 대한 방제효과를 검정하였다. Photorhabdus sp.를 뿌리혹선충, 뿌리썩이선충, 나선선충, 시스트 선충에 처리한 결과 약 90%의 살선충효과를 나타내었다. 실제 뿌리혹선충이 발생한 경북 예천군, 고령군 등의 시설재배 농가에 Photorhabdus sp.를 처리한 결과 약 95%의 밀도 감소효과를 나타내었고 뿌리혹 및 난낭 수축 등의 변화가 관찰되었다. 이는 photorhabdus sp.를 이용하여 식물기생선충, 혹의 형성, 난 부화율 감소 및 식물의 생리작용 활성화, 각종 토양 장애를 최소화하는 선충 방제제 개발 가능성을 제시한다.

An entomopathogenic bacteria, Xenorhabdus nematophila (Xn) and Photorhabdus temperata subsp temperata (Ptt), suppresses insect immune responses and facilitates its symbiotic nematode development in target insect. Benzylideneacetone (BZA), PY, cPY, Ac-FGV, indole, 2-oxindole and 3-(4-hydroxyphenylpropionic) acid (PHPP) were compounds derived from the bacterial. Their immunosuppressive activities have been induced by inhibitory activity against eicosanoid biosynthesis and used to develop an additive to enhance control efficacy of other commercial microbial insecticides. This study investigated any cytotoxicity of their culture broth and bacterial metabolites on Spodoptera exigua hemocyte. When Xn or Ptt (<100 cells per larva) were injected to larval of S. exigua, the bacteria increased in density with incubation time, while the insent hemocyte numbers significantly and the resulting culture broths were sampled for analysis of their cytotoxicity against S. exigua hemocytes. In addition, the sequential culture broth samples were analyzed in active component chemicals using a reverse phase HPLC. Finally, seven bacterial metabolites were analyzed in relative cytotoxicity against S. exigua. These results suggest that BZA is a major cytotoxic compound.

Bacillus thuringiensis (Bt) is a bacterial biopesticide against insect pests, mainly lepidopterans. Spodoptera exigua and Plutella xylostella exhibit significant decreases in Bt susceptibility in late larval instars. To enhance Bt pathogenicity, we used a mixture treatment of Bt and other bacterial metabolites which possessed significant immunosuppressive activities. Mixtures of Bt with culture broths of Xenorhabdus nematophila (Xn) or Photorhabdus temperata ssp. temperata (Ptt) significantly enhanced the Bt pathogenicity against late larval instars. Different ratios of Bt to bacterial culture broth had significant pathogenicities against last instar P. xylostella and S. exigua. Five compounds identified from the bacterial culture broth also enhanced Bt pathogenicity. After determining the optimal ratios, the mixture was applied to cabbage infested by late ins tar P. xylostella or S. exigua in greenhouse conditions. A mixture of Bt and Xn culture broth killed 100% of both insect pests when it was sprayed twice, while Bt alone killed less than 80% or 60% of P. xylostella and S. exigua, respectively. Other Bt mixtures, including Ptt culture broth or bacterial metabolites, also significantly increased pathogenicity in the semi-field assays. These results demonstrated that the Bt mixtures collectively names 'Bt-Plus' can be developed into potent biopesticides to increase the efficacy of Bt.

An entomopathogenic bacterium, Photorhabdus temperata ssp. temperata (Ptt), suppresses insect immune responses and facilitates its symbiotic nematode development in target insects. The immunosuppressive activity of Ptt enhances pathogenicity of various microbial pesticides including Bacillus thuringiensis (Bt). This study was performed to select a cheap and efficient bacterial culture medium for large scale culturing of the bacteria. Relatively cheap industrial bacterial culture media (MY and M2) were compared to two research media, Luria-Bertani (LB) and tryptic soy broth (TSB). In all tested media, a constant initial population of Ptt multiplied and reached a stationary phase at 48 h. However, more bacterial colony densities were detected in LB and TSB at the stationary phase compared to two industrial media. All bacterial culture broth gave significant synergism to Bt pathogenicity against third instars of the diamondback moth, Plutella xylostella. Production of bacterial metabolites extracted by either hexane or ethyl acetate did not show any significant difference in total mass among four culture media. Reverse phase HPLC separated the four bacterial metabolites, which were not much different in quantities among four bacterial culture broths. This study suggests that two industrial bacterial culture media can be used to economically culture Ptt in a large scale.

Two groups of entomopathogenic bacteria, Xenorhabdus and Photorhabdus, are known to suppress insect immune responses by inhibiting eicosanoid biosynthesis. This study used these bacterial culture broths to develop novel biochemical insecticides against the diamondback moth, Plutella xylostella. Though the bacterial culture broths alone showed little insecticidal activity, they significantly enhanced pathogenicity of Bacillus thuringiensis against the fourth instar larvae of P. xylostella. Sterilization of the bacterial culture broth by autoclaving or 0.2 ㎛ membrane filtering did not influence the synergistic effect on the pathogenicity of B. thuringiensis. Three metablites identified in the culture broth of X. nematophila also showed similar synergistic effects. In field test, both entomopathogenic bacterial culture broth also enhanced the control efficacy of B. thuringiensis against P. xylostella.

Oral toxicities of 5 Photorhabdus temperata ssp. temperata (Ptt) strains collected in different regions of Korea were determined against the larvae of Plodia interpunctella, Galleria mellonella, Lucilia caesar, Culex pipiens pallens and Paratlanticus ussuriensis. When a diet or water containing culture media of 5 different Ptt strains were ingested to immature insects, mortalities of the first instar larvae of G. mellonella, L. caesar, P. ussuriensis and young nymphs of C. pipiens pallens were rapidly increased and 100% within 3-5 days after treatments. However, mortality of P. interpunctella neonate larvae was slightly slower and 94.4-100% within 7 days after treatments. As controls, a diet containing either water, the medium without culturing bacteria, or E. coli culture medium did not effective on their mortalities. As another control group, the culture medium of P. temperata ssp. laumondii (KACC) were variously effective to mortalities of 4 species, namely, 100, 45.3, 2.8 and 0% to Galleria, Lucilia, Plodia and Culex, respectively. Culture media of Ptt strains inhibited developmental late of late larvae of P. interpunctella. Our results suggest that the oral administration of the culture medium of Ptt symbiotic bacteria was highly effective to control various immature insects.

Two entomopathogenic bacteria, Xenorhabdus nematophila and Photorhabdus temperata subsp. temperata, are known to be potent against the diamondback moth, Plutella xylostella, when the bacteria are injected into the hemocoel. This study investigated any pathogenic effect of their culture broth on P. xylostella by oral administration. Only culture broth of both bacterial species did not give enough pathogenic effects by the oral administration. However, when the culture broth was orally treated together with Bacillus thuringiensis (Bt), both cell-free culture broth significantly enhanced Bt pathogenicity against the 3rd instar larvae of P. xylostella. The culture broth was then fractionated into hexane, ethyl acetate, and aqueous extracts. Most synergistic effect on Bt pathogenicity was found in ethyl acetate extracts of both bacterial species. Thin layer chromatography of these extracts clearly showed that ethyl acetate extracts of both bacterial culture broths possessed metabolites that were different to those of hexane and aqueous extracts. These results suggest that the both entomopathogenic bacteria produce and secrete different factors to give significant synergistic effect on Bt pathogenicity.

Two entomopathogenic bacteria, Xenorhabdus nematophila (Xn) and Photorhabdus temperata temperata (Ptt), are symbionts of nematodes, Steinernema carpocapsae and Heterorhabditis megidis, respectively. When the nematodes enter host insect hemocoel, the bacteria are released from the nematode intestine to insect hemocoel and cause immunosuppression, which results in septicemia. Culture broth of both bacteria had insecticidal effects when injected into hemocoel of Plutella xylostella larvae, but did not when orally administered. However, either mixture of Xn or Ptt with Bacillus thuringiensis (Bt) significantly enhanced the Bt pathogenicity against P. xylostella. The culture broth was fractionated with hexane and diethylacetate extracts. Diethylacetate extract had potent factor (s) to increase Bt pathogenicity. A compound, benzylideneacetone, identified from the diethylacetate fraction had oral toxicity against P. xylostella. This compound also showed high acaricidal effect on the two spotted spider mite, Tetranychus urticae

Two entomopathogenic bacteria, Xenorhabdus nematophila (Xn) and Photorhabdus temperata temperata (Ptt), maintain monoxenic condition within host insect cadaver by synthesizing and releasing various antibiotics. These two bacteria were cultured in tryptic soy broth during different times, which were screened in their antibacterial activities. Both bacterial culture broth had high antibacterial activities against Escherichia coli at their stationary growth phase. The potent culture broth was used to screen target plant bacterial pathogens using both inhibition zone assay and liquid culture assay. Ralstonia sp. was most susceptible, while Xanthomonas sp. was highly resistant. Pseudomonas sp. and Bacillus sp. showed hemi-susceptible. The culture broth was further fractionated into hexane and diethylether extracts. Significant antibacterial effect was found in the diethylether extract

Two entomopathogenic bacteria, Xenorhadus nematophila and Photorhabdus temperata temperata, are known to suppress immune responses of target insects by inhibiting eicosanoid biosynthesis. This study analyzed these bacterial metabolites in their effects on hemocyte-spreading behavior of the beet armyworm, Spodoptera exigua. Both bacterial culture broth significantly inhibited the hemocyte-spreading behavior, at which the culture broth derived from the stationary growth phase had the most potent effect. Three identified eicosanoid synthesis inhibitors (benzylideneacetone, PY and Ac-FGV) impaired the hemocyte-spreading behavior of S. xigua, at which benzylideneacetone was the most potent. These three compounds share a common chemical structure: a pentenebenzene ring. Alternation of this common structure resulted in significant loss of their inhibitory activity to the hemocyte-spreading behavior.