• BMB Reports
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• 제40권4호
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• pp.494-500
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• 2007

The endophytic bruchid pest Callosobruchus maculatus causes severe damage to storage cowpea seeds, leading to economical losses. For this reason the use of $\alpha$-amylase inhibitors to interfere with the pest digestion process has been an interesting alternative to control bruchids. With this aim, $\alpha$-amylase inhibitors from baru seeds (Dipteryx alata) were isolated by affinity chromatographic procedures, causing enhanced inhibition of C. maculatus and Anthonomus grandis $\alpha$-amylases. To attempt further purification, this fraction was applied onto a reversed-phase HPLC column, generating four peaks with remarkable inhibition toward C. maculatus $\alpha$-amylases. SDS-PAGE and MALDI-ToF analysis identified major proteins of approximately 5.0, 11.0, 20.0 and 55 kDa that showed $\alpha$-amylase inhibition. Results of in vivo bioassays using artificial seeds containing 1.0% (w/w) of baru crude extract revealed 40% cowpea weevil larvae mortality. These results provide evidence that several $\alpha$-amylase inhibitors classes, with biotechnological potential, can be isolated from a single plant species.

• International Journal of Industrial Entomology and Biomaterials
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• 제18권2호
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• pp.125-129
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• 2009

Entomopathogenic fungi were isolated directly from a cadaver of adult Monochamus alternatus supporting fungal sporulation, using a semi-selective medium and then screened several fungal colonies. The pathogenicity of each fungus was tested using oak longicorn beetle, Moechotypa diphysis, as substitutive insect. As the result, only one of them showed high pathogenicity against M. diphysis, with up to 100% mortality within 21 days of inoculation. Selected fungus was named as MaW1 and identified by Beauveria bassiana using microscopic examination and DNA analysis. Pathogenicity was also evaluated to M. alternatus.

• International Journal of Industrial Entomology and Biomaterials
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• 제17권2호
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• pp.211-215
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• 2008

To select entomopathogenic fungi controlling aphids effectively, several isolates were screened against second instars of Myzus persicae nymphs in the glasshouse using conidia suspension at $1.0{\times}10^5$ conidia/ml. Among these isolates, SFB-205 conidia showed the highest insecticidal activity about 32.7% efficacy to M. persicae at 4 days after application in the glasshouse. The attachment of SFB-205 conidia on the surface of M. persicae nymphs, and germination and penetration were observed using scanning electron microscopy. SFB-205 was identified as Beauveria bassiana species through the comparison of 5.8 s rRNA genes. There were 24 polymorphisms between SFB-205 and the previously reported isolate, B. bassiana ATCC74040 using six kinds of primer combinations in amplified fragment length polymorphism (AFLP) analysis. The B. bassiana SFB-205 might be used as a practical biological control agent for the green peach aphid, M. persicae in the field.

• The Plant Pathology Journal
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• 제34권3호
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• pp.250-255
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• 2018

Pine wilt disease, caused by the nematode Bursaphelenchus xylophilus, is one of the most devastating conifer diseases decimating several species of pine trees on a global scale. Here, we report the draft genome of Raoultella ornithinolytica MG, which is isolated from mountain-cultivated ginseng plant as an bacterial endophyte and shows nematicidal activity against B. xylophilus. Our analysis of R. ornithinolytica MG genome showed that it possesses many genes encoding potential nematicidal factors in addition to some secondary metabolite biosynthetic gene clusters that may contribute to the observed nematicidal activity of the strain. Furthermore, the genome was lacking key components of avermectin gene cluster, suggesting that nematicidal activity of the bacterium is not likely due to the famous anthelmintic agent of wide-spread use, avermectin. This genomic information of R. ornithinolytica will provide basis for identification and engineering of genes and their products toward control of pine wilt disease.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.737-744
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• 2004

A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Vairimorpha spp. and Nosema spp. and identification of Vairimorpha necatrix from Lepidoptera insects. Three sets of primers were selected from different genomic sequences to specifically amplify an 831 bp amplicon within the SSU rRNA gene, specific for both Vairimorpha spp. and Nosema spp. (MSSR primer); a 542 bp amplicon within the SSU rRNA gene, specific for Vairimorpha spp. (VSSU primer); and a 476 bp amplicon within the actin gene, specific for Vairimorpha necatrix (VNAG primer). Using the primers in conjunction with multiplex PCR, it was possible to detect Vairimorpha spp. and Nosema spp. and to differentiate between them. The sensitivity of this PCR assay was approximately 10 spores per milliliter. It is proposed that the multiplex PCR is a sensitive, specific, and rapid tool that can serve as a useful differential diagnostic tool for detecting Vairimorpha spp. and Nosema spp. in Lepidoptera insect.

• Journal of Microbiology and Biotechnology
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• 제14권4호
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• pp.772-776
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• 2004

Physiological and molecular characteristics of Bacillus thuringiensis SR6 and SR8 were investigated, and phase contrast and electron microscopies revealed that a large rhomboidal crystal protein was present in the sporulating cells. SDS-PAGE and Western blot analyses showed that B. thuringiensis SR8 produced 70 kDa protein much more than other proteins, and that the 70 kDa protein could bind to the antibody of B. thuringiensis subsp. tenebrionis-crystal toxin protein, indicating that the crystal 70 kDa protein has an immunological homology with B. thuringiensis subsp. tenebrionis-crystal toxin protein. The DNA fragment of B. thuringiensis subsp. tenebrionis-toxin gene was detected in B. thuringiensis SR6 and SR8 by using PCR amplification analysis. Furthermore, the insect bioassay showed the insecticidal activity against Colorado potato beetle larvae. Based on the physiological and molecular similarities to B. thuringiensis subsp. tenebrionis, it is suggested that the B. thuringiensis SR6 and SR8 may be mutants of the B. thuringiensis subsp. tenebrionis strain overexpressing the crystal of 70 kDa toxin protein.

• Journal of Microbiology and Biotechnology
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• 제9권3호
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• pp.334-339
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• 1999

More innovative molecular markers were investigated for rapid and consistent differentiation of Metarhizium anisopliae var. majus from M. anisopliae var. anisopliae. A total of 28 isolates were obtained from various countries and hosts: 13 isolates of M. anisopliae var. anisopliae, 12 isolates of M. anisopliae var. majus, and 3 isolates of M. anisopliae collected in Korea. This study involved restriction enzyme digestions of a PCR product amplified from nuclear internally transcribed spacer (ITS) and a portion of the 28S rDNA regions. Among 11 different restriction enzymes used in this study, MboⅠ digestion particularly produced a restriction pattern that had characteristics of M. anisopliae var. majus. This restriction pattern was consistent in all isolates of M. anisopliae var. majus regardless of their geographic origins and insect hosts. Mapping experiments revealed that MboⅠ sites of M. anisopliae var. majus are identical to those of M. anisopliae var. anisopliae with an exception for the presence of an additional site in the PCR product. Results from this study provide an additional method for identification and differentiation of isolates of these two varieties of M. anisopliae for use in the field and laboratory experiments.

• Animal Systematics, Evolution and Diversity
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• 제31권4호
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• pp.209-246
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• 2015

Morphological features of larvae and pupae of twelve species of Simulium (Simulium) from Korea are investigated and redescribed. Detailed descriptions of immature stages of each species are provided with photographs. For the Korean members of subgenus Simulium, combinations of the following characteristics can be used to separate them from other Korean subgenera: larval abdomen without a pair of conical-shaped ventral tubercles; postgenal cleft variously shaped, viz., miter shaped, spearhead, bullet shaped or triangular; rectal papillae 3 simple lobe or compound lobes; pupal gill of 6-16 filaments; cocoon shoe, boot or slipper shaped; and head trichomes 3 pairs (1 facial+2 frontal). Habitat information and other ecological aspects of each species are provided. Updated keys to larvae and pupae of the Korean species of the subgenus Simulium are also provided to facilitate species identification. Taxonomic treatment based on external morphologies of larvae and pupae of Korean species of the subgenus Simulium are focused and included in this study. To understand their interspecific relationships, phylogenetic analyses using multiple data from molecule, morphology, and ecology are warranted in future research. Also continued exploration for new character systems is needed to establish more stable boundaries for subgenus and species delimitations.

• Proceedings of the National Institute of Ecology of the Republic of Korea
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• 제1권1호
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• pp.52-57
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• 2020

Bacterial symbionts are common across insects, including ants (Hymenoptera: Formicidae). Reproduction-manipulating endosymbionts, such as Wolbachia, Spiroplasma, Rickettsia, and Cardinium, are closely associated with many aspects of host-insect life. In addition, phage WO plays an essential role in the phenotypic effects of Wolbachia. Although endosymbionts are possible biological control agents, there is a lack of knowledge of their rate of infection of ants in Korea. We tested a range of Korean ant species for the presence of Wolbachia, Spiroplasma, Rickettsia, Cardinium, and phage WO by extracting DNA from the ants and using specific primer sets to test the status of infections. In addition, the mitochondrial cytochrome c oxidase I (COI) gene of the host ants was amplified to confirm the molecular identification and phylogenetic relationship between the hosts. We found that infection with Wolbachia (29.6% of species) is relatively common when compared with that of other endosymbionts. Only one species was infected with Spiroplasma. Infection with Rickettsia and Cardinium was not detected in the examined ants. Most Wolbachia in ants were infected with phage WO. Although the phenotypic effects of endosymbionts in ants are still unknown, this first survey of endosymbionts in Korea is the first step toward the use of reproduction-manipulating endosymbionts.

• Journal of Microbiology and Biotechnology
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• 제33권6호
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• pp.745-752
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• 2023

Gut symbionts play crucial roles in host development by producing nutrients and defending against pathogens. Phloem-feeding insects in particular lack essential nutrients in their diets, and thus, gut symbionts are required for their development. Gram-negative Pantoea spp. are known to be symbiotic to the western flower thrips (Frankliniella occidentalis). However, their bacterial characteristics have not been thoroughly investigated. In this study, we isolated three different bacteria (BFoK1, BFiK1, and BTtK1) from F. occidentalis, F. intonsa, and T. tabaci. The bacterial isolates of all three species contained Pantoea spp. Their 16S rRNA sequences indicated that BFoK1 and BTtK1 were similar to P. agglomerans, while BFiK1 was similar to P. dispersa. These predictions were supported by the biochemical characteristics assessed by fatty acid composition and organic carbon utilization. In the bacterial morphological analysis, BFoK1 and BTtK1 were distinct from BFiK1. All these bacteria were relatively resistant to tetracycline compared to ampicillin and kanamycin, in which BFoK1 and BTtK1 were different from BFiK1. Feeding ampicillin (100,000 ppm) reduced the bacterial density in thrips and retarded the development of F. occidentalis. The addition of BFoK1 bacteria, however, rescued the retarded development. These findings indicate that Pantoea bacteria are symbionts to different species of thrips.