Spodoptera eridania and S. ornithogalli (Lepidoptera: Noctuidae), which are polyphagous pests that damage various crops such as tomatoes and beans are regulated quarantine species that are highly likely to invade South Korea. Therefore, it is crucial to promptly and accurately identify the presence of S. eridania and S. ornithogalli in crop fields to effectively eradicate as a regulated quarantine species. In this study, we developed a loop-mediated isothermal amplification (LAMP) assay, which allows for rapid in-field identification. To develop the LAMP　assay, we selected target species-specific genomic regions from the whole-genome sequences of one target and 13 other lepidopteran species. We validated each five and six primer sets that consistently produced positive reactions in S. eridania and S. ornithogalli, respectively. To test the sensitivity of the each locus, LAMP reactions were performed using various reaction times using crude DNA, which was extracted from various types of adult tissues. All sensitivity tests were also successful.

The purpose of this study is to develop the quantitative PCR(qPCR) assay that would enable the rapid identification and simultaneous detection of six different endodontic pathogenic bacteria in a single reaction. In this study, six pairs of primers for Treponema denticola, Porphyromonas gingivalis, Fusobacterium nucleatum, Prevotella intermedia, Streptococcus mutans, and Staphylococcus aureus and two pairs of housekeeping genes were designed for a multiplex qPCR based on the SYBR Green method. The genomic DNA was extracted from reference strains and submitted to the qPCR reaction. The specificity of the amplified products was analyzed by melting curves. As a result, six distinct melting peaks were identified by the melting curve analysis and all of the target species were simultaneously discriminated. Therefore, the multiplex qPCR assay developed in this study can be used for rapid identification and detection of T. denticola, P. gingivalis, F. nucleatum, P. intermedia, S. mutans, and S. aureus at the same time. In combination with the melting curve analysis, the level of the target species and total bacterial load can be obtained.

Insect resistance to crop protecting chemicals is developing very rapidly, which is a major problem in the production of crops. Recently, the mechanism of resistance to various pseticides s has been revealed in Helicoverpa armigera. However, to date, no technology has been developed to quickly diagnose resistance yet. In this study, we present the results of developing method of rapid diagnosis of resistance

The tobacco budworm, Helicoverpa assulta, is a freeze-susceptible species that overwinters in temperate zones with pupa diapause. A rapid cold hardening (RCH) and supercooling capacity usually play crucial roles in survival during the overwintering period. This study is performed to identify a cryoprotectant as a RCH factor in H. assulta. Pre-exposure of H. assulta larvae to 4°C significantly increased survival at -10°C in all developmental stages from egg to adult. RCH was dependent on the duration of the pre-exposure period. RCH also significantly enhanced the supercooling capacity. Cryoprotectant analysis using HPLC showed that the preexposure treatment allowed the larvae to accumulate glycerol in the hemolymph. Two genes, glycerol-3-phosphate dehydrogenase (GPDH) and glycerol 3-phosphatase (G3P), that involing in glycerol biosynthesis were identified from the transcriptome of H. assulta 4th instar larvae. From the result of transcriptome, the expressions of GPDH and G3P were relatively increased when compared to that of the control, suggesting that these genes contribute to overwintering and biosynthesis of cryoprotectant.

A positional scanning synthetic peptide combinatorial library (PS-SCL) was screened in order to identify antimicrobial peptides against the cariogenic oral bacteria, Streptococcus mutans. Activity against Streptococcus gordonii and Aggregatibacter actinomycetemcomitans was also examined. The library was comprised of six sub-libraries with the format O(1-6)XXXXX-NH2, where O represents one of 19 amino acids (excluding cysteine) and X represents equimolar mixture of these. Each sub-library was tested for antimicrobial activity against S. mutans and evaluated for antimicrobial activity against S. gordonii and A. actinomycetemcomitans. The effect of peptides was observed using transmission electron microscopy (TEM). Two semi-mixture peptides, RXXXXN-NH2 (pep-1) and WXXXXN-NH2 (pep-2), and one positioned peptide, RRRWRN-NH2 (pep-3), were identified. Pep-1 and pep-2 showed significant antimicrobial activity against Gram positive bacteria (S. mutans and S. gordonii), but not against Gram negative bacteria (A. actinomycetemcomitans). However, pep-3 showed very low antimicrobial activity against all three bacteria. Pep-3 did not form an amphiphilic α-helix, which is a required structure for most antimicrobial peptides. Pep-1 and pep-2 were able to disrupt the membrane of S. mutans. Small libraries of biochemically-constrained peptides can be used to generate antimicrobial peptides against S. mutans and other oral microbes. Peptides derived from such libraries may be candidate antimicrobial agents for the treatment of oral microorganisms.

벼 흰잎마름병균의 정확한 진단을 위하여 PCR용 진단 kit를 개발하였다. 본 PCR kit를 개발하기 위하여 벼 흰잎마름 병균 유전체 정보 중 phage-related integrase and transposase gene의 염기서열을 이용하여 프라이머를 각각 제작하였다. 프라이머 염기서열은 XOP-F (5-CGG TCT GCT CAA TGA GGA AGA-3)와 XOP-R2 (5-TGC AAT TGG TGT TCTCCA GG-3), XOT-F (5-GTC ATA GGT GAG GCT TCCC-3)와 XOT-R2 (5-AGT GCG ATC TTT CAG CAG G-3)로 벼 흰잎마름병균의 DNA를 401bp와 492bp를 증폭하게 제작하였다. PCR 증폭은 벼 흰잎마름병균만 증폭하였으며 다른 세균인 Escherichia coli, Agrobacterim, Pectobacterium caratovora subsp. cartovorum, P. atrosepticum, Pseudomonasputida, P. syringae, P. savastanoi pv. phaeolicola, P. savastanoipv. savastanoi and P. marginalis pv. Marginalis 등은증폭되지 않아 특이성이 인정 되었다. 본 프라이머로 병이 의심되는 벼잎과 논물에서 병원균을 3시간 이내에 검출할 수 있었다.

The infectious pathogens against honeybee (Apis mellifera) comprise a heterogeneous group of bacterial, viral, and fungal organisms including Paenibacillus larvae, Deformed Wing Virus (DWV) and Nosema apis. Many species like Paenibacillus larvae, Deformed Wing Virus (DWV) and Nosema apis have been isolated from a number of different continents, e.g. America, Asia and Europe, indicating its wide spread in whole nature. Little is known about the occurrence and distribution in the environment of these pathogens. For a more rapid, systematic and efficient monitoring of each pathogenic species against honeybee in the environment, PCR-based detection systems were developed that allows species-specific identifications of various pathogenic species with one reaction. These could be achieved by selecting specific primers from conserved regions of each species with speciesspecific DNA sequence variations. For the detection of any already known pathogen, well-developed PCR-detection system allows the specific detection of expected pathogenic species based on its specific nucleic acid sequence. Since each pathogenic species delivers a specific PCR-product of different size, bands can be distinguished very easily by simple gel electrophoresis. After the development of real-time PCR system, PCR-based specific detections of honeybee pathogens were dramatically improved their applications, from just detection to quantification of pathogens. These systems, quantitative PCR (qPCR) for the detection of honeybee pathogens, could be distinguished from previous PCR detection on the points of “real-time”, “easy” and “quantitative”. Moreover, very rapid PCR, so-called “Ultra-Rapid Real-Time PCR” were developed recently in field of pathogen-detection. Typical Honeybee pathogens such as Paenibacillus larvae, Israelli acute paralysis virus (IAPV) were successfully detected inner 7 minutes using 30 cycled Ultrarapid PCR. According to development of more rapid apparatus, even 30 cycled, 1 minute PCR seems to be possible. Ultla-Rapid PCR was currently attempted to apply for the direct detection system of all viral pathogens against honeybee from bee-samples and different environmental probes.

This study describes the efficient method for the discrimination of 'Cheonryang' in Panax ginseng Meyer using a STS primer. A total of 208 STS primers were applied to polymerase chain reaction (PCR) amplification for discriminating Korean ginseng cultivars. Co-dominant polymorphic band patterns were generated with two primers, MFGp 0019, MFGp 0248, and successful identification of 'Cheonryang' was achieved from out of 11 Korean ginseng cultivars. Two different sizes of DNA band patterns were detected with MFGp 0019 primer. Ten Korean ginseng cultivars shared the same size of amplified DNAs (389 bp), but 'Cheonryang' showed a different size. Thus 'Cheonryang' can be efficiently distinguished from the other ten ginseng cultivars by using the MFGp 0019 primer. In the case of MFGp 0248, two different sizes of DNA band patterns were detected in the eleven ginseng cultivars. Same sized amplified DNA bands (307 bp) were shown in five cultivars (Chunpoong, Gopoong, Kumpoong, Cheongsun, Sunhyang) and 254 bp sized DNA bands were identified in the other 6 cultivars (Yunpoong, Sunpoong, Sunun, Sunone, Cheonryang, K-1). In conclusion, the two STS primers, MFGp 0019, and MFGp 0248, provide a rapid and reliable method for the specific identification of 'Cheonryang' cultivar from a large number of samples.

Receptor mediated signal carriers play a critical role in regulation of plant defense and development. Rapid Alkalization Factor (RALF) is an important signaling family which has a role in plant growth and development. However, only few RALF polypeptides have been identified till date, mainly because of enormous efforts required for their isolation or identify their gene through mutational analysis. In this study, an extensive database search yield 39, 43, 34 and 23 potential RALF genes in Arabidopsis, rice, corn and soybeans, respectively. RALF genes are highly conserved across the plant species. A comprehensive analysis including the chromosomal location, gene structure, subcellular location, conserved motif, protein structure and promoter analysis was performed. RALF genes from four plants under study were divided in 7 groups based on phylogenetic analysis. In silico expression analysis of these genes, using microarray and EST data, reveled that these genes exhibit a variety of expression pattern. Furthermore, RALF genes showed distinct expression pattern under nitricoxide (NO) stress in Arabidopsis. This suggests a role of RALF genes in plant defense regulation. Our comprehensive analysis of RALF genes is a valuable resource that further elucidates the roles of RALF family members in plant growth and development. In addition, comparative genomics analyses deepen our understanding of the evolution of RALF gene family and will contribute to further genetics and genomics studies of other monocot and dicot plant species.