• 식물병연구
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• 제29권2호
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• pp.130-136
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• 2023

Citrus melanoses caused by Diaporthe citri, has been one of the serious diseases in many citrus orchards of Jeju Island. To protect melanose in citrus farms, a fast and exact diagnosis method is necessary. In this study, diseased leaves and dieback twigs were collected from a total of 49 farms within March to April in 2022. A total of 465 fungal isolates were obtained from a total of 358 isolated plant samples. Among these fungal isolates, 40 representatives of D. citri isolates which were isolated from 22 twigs and 18 leaves on 23 farms were found based on cultural characteristics on potato dextrose agar and conidial morphology. Additionally, the molecular assay was carried out and compared with those by morphological diagnosis. All isolates were identified as D. citri by analyzing the sequences at the internal transcribed spacer (ITS) rDNA region using primers of ITS1/ITS4 or at β-tubulin using primer Btdcitri-F/R. Therefore, based on the present study, where the results of morphological identification of conidial type were consistent with DNA sequence analysis of certain gene, choosing a suitable method for a fast diagnosis of citrus melanose was suggested.

• Mycobiology
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• 제35권4호
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• pp.171-173
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• 2007

Some Xylaria materials growing on the fruits of Liquidambar spp. were collected. They were identified as X. persicaria on the basis of morphological characteristics and sequence analysis of the complete ITS region (ITS1-5.8S-ITS2) of rDNA. This is the first record of this species from Korea.

• Animal cells and systems
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• 제15권4호
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• pp.315-324
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• 2011

The actiniarian sea anemone, Entacmaea quadricolor, and the scleractinian coral, Alveopora japonica, host symbiotic dinoflagellates belonging to the genus Symbiodinium (Freudenthal). We studied the host-symbiont specificity of these two anthozoan hosts in the northwestern Pacific Ocean. Symbionts within the two hosts were identified using partial large subunit (LSU) ribosomal DNA (rDNA) and complete internal transcribed spacers (ITS) 1 rDNA regions. The host, E. quadricolor, was identified using the partial LSU rDNA molecular marker. Genetic analysis showed that E. quadricolor only harbors dinoflagellates belonging to subclade C1/3 of the genus Symbiodinium. Moreover, no genetic variation was detected among the symbionts of E. quadricolor within the study region (Korea and Japan), even though the two distant sites were separated by more than 1000 km, at collection depths of 1 m in shallow and 13-16 m in deep water. Whilst scleractinian corals host multiple Symbiodinium clades in tropical waters, A. japonica, sampled over a wide geographical range (800 km) within the study region, only hosts Symbiodinium sp. clade F3. The high specificity of endosymbionts in E. quadricolor and A. japonica within the northwestern Pacific Ocean could be accounted for because symbiotic dinoflagellates within the host anemones appear to be acquired maternally, and the Kuroshio Current might affect the marine biota of the northwestern Pacific. However, the consistency of the symbiotic relationships between these two anthozoan hosts and their endosymbionts could change after climate change, so this symbiotic specificity should be monitored.

• IMMUNE NETWORK
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• 제11권5호
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• pp.268-280
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• 2011

Background: Dengue virus, which belongs to the Flavivirus genus of the Flaviviridae family, causes fatal dengue hemorrhagic fever (DHF) and dengue shock syndrome (DSS) with infection risk of 2.5 billion people worldwide. However, approved vaccines are still not available. Here, we explored the immune responses induced by alternating prime-boost vaccination using DNA vaccine, adenovirus, and vaccinia virus expressing E protein of dengue virus type 2 (DenV2). Methods: Following immunization with DNA vaccine (pDE), adenovirus (rAd-E), and/or vaccinia virus (VV-E) expressing E protein, E protein-specific IgG and its isotypes were determined by conventional ELISA. Intracellular CD154 and cytokine staining was used for enumerating CD4+ T cells specific for E protein. E protein-specific CD8+ T cell responses were evaluated by in vivo CTL killing activity and intracellular IFN-${\gamma}$ staining. Results: Among three constructs, VV-E induced the most potent IgG responses, Th1-type cytokine production by stimulated CD4+ T cells, and the CD8+ T cell response. Furthermore, when the three constructs were used for alternating prime-boost vaccination, the results revealed a different pattern of CD4+ and CD8+ T cell responses. i) Priming with VV-E induced higher E-specific IgG level but it was decreased rapidly. ii) Strong CD8+ T cell responses specific for E protein were induced when VV-E was used for the priming step, and such CD8+ T cell responses were significantly boosted with pDE. iii) Priming with rAd-E induced stronger CD4+ T cell responses which subsequently boosted with pDE to a greater extent than VV-E and rAd-E. Conclusion: These results indicate that priming with live viral vector vaccines could induce different patterns of E protein-specific CD4+ and CD8+ T cell responses which were significantly enhanced by booster vaccination with the DNA vaccine. Therefore, our observation will provide valuable information for the establishment of optimal prime-boost vaccination against DenV.

• Journal of Animal Science and Technology
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• 제47권3호
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• pp.465-474
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• 2005

A two-step broad-range PCR method detecting gram-negative bacteria at the level as low as 2 CFU was developed by using primers of GNFI and GNRI and then semi-nested primer of GNF2 and GNRI. The nucleotide sequences of the primers were determined based on l6S rRNA gene. The DNA fragments of 1173 bp and 169 bp were amplified in one-step PCRs with primer sets of GNFI-GNRI and GNF2-GNRl, respectively, using template DNA from seven strains of gram-negative bacteria including Escherichia coli, Enterobacter aerogenes, Klebsiella pneumoniae, Pseudomonas spp., and Acinetobacter baumaii but not from Achromobacter lyticus, Alca/igens faecalis, and five strains of gram-positive bacteria. DNA fragments of 180 bp were amplified from LTLT-pasteurized milk and UHf-pasteurized milk in the two-step PCR. The DNA fragments were amplified from LTLT-pasteurized milk which was added with Pseudomonas j/uorescens and subsequently heated at 65 $^{\circ}C$, 80 $^{\circ}C$, and 100 $^{\circ}C$ for 30 min but they were not amplified from the milk autoclaved at 121$^{\circ}C$ for 15 min. It was suggested in PCR that Pseudomonas fluorescens heated at 65 $^{\circ}C$ for 30 min in milk was more sensitive to DNase treatment than viable bacteria.

• 미생물학회지
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• 제32권3호
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• pp.186-191
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• 1994

Staphylococcus aureus로부터 분리한 R-plasmid pSBK203의 복제개시 단백질인 Rep의 작용부위인 ori 및 dsDNA로의 전환을 위해 요구되는 minus origin부위를 밝히고자 시도하였다. Escherichia coli vecotr를 이용하여 pSBK203의 복제관련 부위를 최소한도로 포함하는 재조합 E.coli-Bacillus subtilis shuttle vector를 구성, 분리하고 여기에 포함된 pSBK203부위의 염기 서열을 분석함으로써 ori를 확인하였다. pSBK203의 복제개시 부위 ori는 rep의 구조 유전자 ORF내에서 약 50bp의 크기로 발견되었으며 지금까지 알려진 staphylococcal plasmid들중에서 pT181족 plasmid들의 ori와 높은 상동성을 갖는 것으로 분석되었다. 복제 과정에서 ssDNA로 먼저 만들어진 (+)쇄가 dsDNA로 전환되기 위해 필요한 신호로 작용하는 것으로 알려저 있는 minus origin (M-O)인 긴 palindrome 구조, 즉 pal 부위가 rep 우전자의 상류에서 2개 연이어 존재하는 것이 발견되었다. 이중에서 pOX6, pC194, 및 pE194 등과 같은 다른 staphylococcal plasmid들의 pal 부위와 비교적 높은 상동성을 갖는 paLA 는 plasmid 유지에 별 영향을 미치지 못하는 반면 다른 plasmid에서 유사 서열이 보고되지 않은 palA는 plasmid 유지에 필수적이라는 사실이 밝혀졌다.

• 생명과학회지
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• 제16권1호
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• pp.71-75
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• 2006

We analyzed the phylogenic relationships of 23 partial 18S rDNA sequences of 22 species (1 species has 2 strains) belonging to Sarcorptiforms include 4 new sequences, using several tools. Although geographic distributions are quite far from, sequence similarity of two strains of Dermatophygoides pteronyssinus isolated from Japan and New Zealand were very high. This result suggests that mite migration by animals including human occurred in the two continents. We investigated the Endeostigmata taxonomic relationship between the Prostigmata and Oribatida subgroups using small fragments (340-400 bp) of their 185 rDNA sequences. But Endeostigmata was not grouped with Oribatida or Prostigmata. In conclusion, it is first reported phylogenic relationship for classified mites included in Sarcoptiformes using 185 rDNA sequence analysis and its system is a very powerful tool for classification of mites.

• Journal of Microbiology and Biotechnology
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• 제16권1호
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• pp.68-76
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• 2006

Despites many attempts to explore the microbial diversity in kimchi fermentation, the predominant flora remains controversial to date. In the present study, major lactic acid bacteria (LAB) were investigated in Chinese cabbage kimchi in the early phase of fermention. For the samples over pH 4.0, viable cell counts of Leuconostoc and Pediococcus were $10^6\;cfu/ml$ and below $10^2\;cfu/ml$, respectively, and 20 isolates out of 172 were subjected to a biochemical identification (API 50 CH kit) as well as molecular-typing methods including ITSPCR with a RsaI digestion and 16s rRNA gene sequence analysis for species confirmation. Seven isolates were nicely assigned to Lb. brevis, 6 to Leuconostoc spp. (2 mesenteroides, 2 citreum, I carnosum, I gasicomitatum), 4 to Weissella (3 kimchii/cibaria, 1 hanii) and 2 to other Lactobacillus spp. (1 farciminis, 1 plantarum). On the other hand, the biochemical identification data revealed 9 strains of Lb. brevis, 6 strains of Leuconostocs,2 strains of Lb. plantarum and 1 strain each of Lb. coprophilus and Lactococcus lactis. However, a single isolates, YSM 16, was not matched to the ITS-PCR database constructed in the present study. Two Lb. brevis strains by API 50 CH kit were reassigned to W kimchii/cibaria, Lb. coprophilus or W hanii, respectively, judging from the results by the above molecular typing approaches. As a whole, the identification data obtained by the biochemical test were different from those of ITS-PCR molecular method by about $63\%$ at genus-level and $42\%$ at species-level. The data by the ITS-PCR method conclusively suggest that predominant LAB species is probably heterolactic Lb. brevis, followed by W kimchii/cibaria, Leuc. mesenteroides, and Leuc. citreum, in contrast to the previous reports [3] that Leuc. mesenteroides is the only a predominant species in the early phase kimchi fermentation.

• 한국응용곤충학회지
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• 제53권3호
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• pp.247-260
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• 2014

옥수수 포장에서 발생하는 Trichogramma속 조명나방 알기생벌의 종 분포를 조사하기 위해 채집된 알기생벌로부터 핵내 ITS2 DNA 전체 염기서열 정보를 해독하였다. 그리고 종 구별을 위한 참고정보로 NCBI GenBank에 등록된 Trichogramma속 60종의 ITS2 전체 염기서열을 확보하여 비교하였다. 국내 채집 알기생벌은 ITS2 DNA 길이와 3' 말단 염기서열 패턴에 따라 3개 그룹(K-1, -2, -3)으로 구분되었다. 국내 채집 그룹 내 염기서열 차이 추정값(Evolutionary distance, d)은 0.005 이하로 그룹 간 비교 시 d 값(${\geq}0.080$)보다 낮았다. 그룹 및 GenBank 등록 종 간 비교시 K-1은 T. ostriniae, K-2는 T. dendrolimi, K-3은 T. confusum과 d 값이 각각 0.016, 0.001, 0.002로 가장 작았다. 추론된 분자계통수에서 K-1은 T. ostriniae, K-2는 T. dendrolimi와 각각 분지되었으나 K-3는 T. confusum, T. chilonis, T. bilingensis와 함께 분지되었다. NCBI BLAST 결과에서도 K-1은 T. ostriniae와 K-2는 T. dendrolimi와 99% identity를 보였다. 그러나 K-3의 홍천 채집 기생벌들은 T. confusum, T. chilonis와 99-100% identity를 보였지만, 고창 채집 기생벌은 T. bilingensis, T. confusum, T. chilonis와 98% identity를 보였다. 이상의 분석 결과 본 연구에서 채집된 알기생벌 K-1과 K-2는 각각 T. ostriniae와 T. dendrolimi, 단일한 종으로 추정되었으나 K-3는 ITS2 정보만으로 종을 추정하기 어려웠다.

• Food Science and Biotechnology
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• 제14권3호
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• pp.416-419
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• 2005

Unknown bacterium isolated from garlic was characterized using phenotypic methods, phylogenetic analysis, DNA-DNA hybridization, and cultural methods. The strain was identified as typical leuconostoc; Gram-positive, non-sporeforming, heterofermentative, catalase-negative and spherical. Although its 16S rRNA gene sequence showed high homology to Leuconostoc argentinum DSM $8581^T$(99.8%), DNA-DNA hybridization experiments indicated it represents novel genomic species in the genus Leuconostoc. The garlic-specific leuconostoc was more resistant to antimicrobial activity of garlic compared to other common laboratory lactic acid bacteria, and was even stimulated by low concentrations (1-2%) of garlic extract supplemented in trypticase soy broth. Growth stimulation was concentration-dependent when tested with residual aqueous layer after solvent extraction of fresh whole garlic extract.