• Mycobiology
• /
• 제33권2호
• /
• pp.104-108
• /
• 2005

Genus Phellinus taxonomically belongs to Aphyllophorales and some species of this genus have been used as a medicinal ingredients and Indian folk medicines. Especially, P. linteus and morphological-related species are well-known medicinal fungi that have various biological activities such as humoral and cell-mediated, anti-mutagenic, and anti-cancer activities. However, little is known about the rapid detection for complex Phellinus species. Therefore, this study was carried out to develop specific primers for the rapid detection of P. linteus and other related species. Designing the species-specific primers was done based on internal transcribed spacer sequence data. Each primer set detected specifically P. linteus (PL2/PL5R) and P. baumii (PB1/PB4R). These primer sets could be useful for the rapid detection of specific-species among unidentified Phellinus species. Moreover, restriction fragment length polymorphism analysis of the ITS region with HaeIII was also useful for clarifying the relationship between each 5 Phellinus species.

• 한국미생물학회:학술대회논문집
• /
• 한국미생물학회 2002년도 추계학술대회
• /
• pp.100-105
• /
• 2002

The diversity of epiphytic bacterial communities on deciduous oak tree (Quercus dentate Thunb.) leaves was examined both in the natural forest area with a clean air and in the industrial estate to assess effects of acidic deposition to the phyllosphere using 16S rDNA sequence data. In addition, acid-tolerant epiphytic bacterial communities were compared. A total of 78 epiphytic and 444 acid-tolerant clones were obtained from clone libraries, resulting in 20 and 17 phylotypes by analysis of restriction fragment length polymorphism (RFLP) for PCR-amplified 16S rDNA products. A low bacterial diversity in both areas was found. As tree leaves grow older, bacterial diversities were slightly increased in the level of subphylum. The community structure of epiphytic bacteria in both areas in April consisted of only two subphyla, $\beta-and\;\gamma-Proteobacteria$. In August two additional subphyla in both areas were found, but the composition was a little different, Acidobacteria and Cytophaga-Flexibacter-Bacteroids (CFB) group in the industrial estate and a -Proteobacteria and CFB group in the natural area, respectively. Acidobacteria could be an indicator of epiphytic bacteria for acidic deposition on plant leaves, whereas a -Proteobacteria be one of epiphytic bacteria that naturally survive on leaves that are not affected by acidic deposition. The acid-tolerant bacterial communities in April were composed of two subphyla, $\gamma-Proteobacteria$ and Low G+C gram-positive bacteria in both areas, and in August a-Proteobacteria was added to the community just in the natural forest area. The direct influence of acidic deposition on the acid-tolerant bacterial phylogenetic composition could not be detected in higher taxonomic levels such as subphylum, but at narrower or finer levels it could be observed by a detection of Xanthomonadales group of $\gamma-Proteobacteria$ just in the industrial estate.

• Journal of Animal Science and Technology
• /
• 제55권2호
• /
• pp.87-93
• /
• 2013

We identified four new bovine tri-nucleotide microsatellite loci and analyzed their sequence structures and genetic parameters in 105 randomly selected Korean cattle (Hanwoo). Allele numbers of the loci B17S0808, B15S6253, B8S7996, and B17S4998 were 10, 11, 12, and 29, respectively. These alleles contained a simple or compound repeat sequences with some variations. Allele distributions of all these loci were in Hardy-Weinberg equilibrium (P > 0.05). Observed heterozygosity and expected heterozygosity ranged from 0.54 (B15S6253) to 0.92 (B17S4998) and from 0.599 (B15S6253) to 0.968 (B17S4998), respectively, and two measures of heterozygosity at each locus were highly correlated. Polymorphism information content (PIC) for these 4 loci ranged from 0.551 (B15S6253) to 0.932 (B17S4998), which means that all these loci are highly informative (PIC > 0.5). Other genetic parameters, power of discrimination (PD) and probability of exclusion (PE) ranged from 0.783 (B15S6253) to 0.984 (B17S4998) and from 0.210 (B15S6253) to 0.782 (B17S4998), respectively. Their combined PD and PE values were 0.9999968 and 0.98005176, respectively. Capillary electrophoresis revealed that average peak height ratio for a stutter was 13.89% at B17S0808, 26.67% at B15S6253, 9.09% at B8S7996, and 43.75% at B17S4998. Although the degree of genetic variability of the locus B15S6253 was relatively low among these four microsatellite markers, their favorable parameters and low peak height ratios for stutters indicate that these four new tri-nucleotide microsatellite loci could be useful multiplex PCR markers for the forensic and population genetic studies in cattle including Korean native breed.

• Parasites, Hosts and Diseases
• /
• 제53권4호
• /
• pp.413-419
• /
• 2015

The present study determined and compared the genetic diversity of Plasmodium falciparum strains infecting children living in 2 areas from Gabon with different malaria endemicity. Blood samples were collected from febrile children from 2008 to 2009 in 2 health centres from rural (Oyem) and urban (Owendo) areas. Genetic diversity was determined in P. falciparum isolates by analyzing the merozoite surface protein-1 (msp1) gene polymorphism using nested-PCR. Overall, 168 children with mild falciparum malaria were included. K1, Ro33, and Mad20 alleles were found in 110 (65.5%), 94 (55.9%), and 35 (20.8%) isolates, respectively, without difference according to the site (P>0.05). Allelic families' frequencies were comparable between children less than 5 years old from the 2 sites; while among the older children the proportions of Ro33 and Mad20 alleles were 1.7 to 2.0 fold higher at Oyem. Thirty-three different alleles were detected, 16 (48.5%) were common to both sites, and 10 out of the 17 specific alleles were found at Oyem. Furthermore, multiple infection carriers were frequent at Oyem (57.7% vs 42.2% at Owendo; P=0.04) where the complexity of infection was of 1.88 (${\pm}0.95$) higher compared to that found at Owendo ($1.55{\pm}0.75$). Extended genetic diversity of P. falciparum strains infecting Gabonese symptomatic children and high multiplicity of infections were observed in rural area. Alleles common to the 2 sites were frequent; the site-specific alleles predominated in the rural area. Such distribution of the alleles should be taken into accounts when designing MSP1 or MSP2 malaria vaccine.

• The Plant Pathology Journal
• /
• 제29권2호
• /
• pp.125-135
• /
• 2013

In agro-ecosystems worldwide, some of the most important and devastating diseases are caused by soil-borne necrotrophic fungal pathogens, against which crop plants generally lack genetic resistance. However, plants have evolved approaches to protect themselves against pathogens by stimulating and supporting specific groups of beneficial microorganisms that have the ability to protect either by direct inhibition of the pathogen or by inducing resistance mechanisms in the plant. One of the best examples of protection of plant roots by antagonistic microbes occurs in soils that are suppressive to take-all disease of wheat. Take-all, caused by Gaeumannomyces graminis var. tritici, is the most economically important root disease of wheat worldwide. Take-all decline (TAD) is the spontaneous decline in incidence and severity of disease after a severe outbreak of take-all during continuous wheat or barley monoculture. TAD occurs worldwide, and in the United States and The Netherlands it results from a build-up of populations of 2,4-diacetylphloroglucinol (2,4-DAPG)-producing fluorescent Pseudomonas spp. during wheat monoculture. The antibiotic 2,4-DAPG has a broad spectrum of activity and is especially active against the take-all pathogen. Based on genotype analysis by repetitive sequence-based-PCR analysis and restriction fragment length polymorphism of phlD, a key 2,4-DAPG biosynthesis gene, at least 22 genotypes of 2,4-DAPG producing fluorescent Pseudomonas spp. have been described worldwide. In this review, we provide an overview of G. graminis var. tritici, the take-all disease, Pseudomonas biocontrol agents, and mechanism of disease suppression.

• 한국작물학회:학술대회논문집
• /
• 한국작물학회 1999년도 춘계 학술대회지
• /
• pp.162-163
• /
• 1999

The goal of this research was (1) to describe the conditions and parameters required for the cell cycle synchronization and the accumulation of large number of metaphase cells in maize and other cereal root tips, (2) to isolate intact metaphase chromosomes from root tips suitable for characterization by flow cytometry, and (3) to construct chromosome-specific libraries from maize. Plant metaphase chromosomes have been successfully synchronized and isolated from many cereal root-tips. DNA synthesis inhibitor (hydroxyurea) was used to synchronize cell cycle, follwed by treatement with trifluralin to accumulate metaphase chromosomes. Maize flow karyotypes show substantial variation among inbred lines. thish variation should be sueful in isolating individual chromosome types. In addition, flow cytometry is a useful method to measure DNA content of individual chromosomes in a genotyps, and to detect chromosomal variations. Individual chromosome peaks have been sorted from the maize hybrid B73/Mol7. Libraries were generated form the DOP-PCR amplification product from each peak. To date, we have analyzed clones from a library constructed from the maize chromosome 1 peak. Hybridization of labeled genomic DNA to clone inserts indicated that $24\%,\;18\%,\;and\;58\%$ of the clones were highly repetitive, medium repetitive, and low copy, respectively. Fifty percent of putative low cpoy clones showed single bands on inbred screening, blots, and the remaining $50\%$ were low copy repeats. Single copy clones showing polymorphism will be mapped using recombinant inbred mapping populations. Repetitive clones are being characterized by Southern blot analysis, and will be screened by in situ hybridization for their potential utility as chromosome specific markers.

• 생명과학회지
• /
• 제21권3호
• /
• pp.468-472
• /
• 2011

DNA 마커는 유전현상 분석이나 품종육성에 널리 사용되고 있다. 본 연구에서는 내냉성 오이 계통인 'NC76'과 냉해 감수성 계통인 'GY14'로부터 내냉성 연관 마커을 목적으로 기존의 총 995개 SSR 마커의 다형성을 평가하였다. Agarose gel 전기영동법으로 'NC76과 'GY14' 간 PCR증폭 산물의 길이 다형성을 보이는 145개 SSR 마커를 개발하였으며, high resolution melting (HRM) 기술을 사용하여 염기서열 다형성을 보이는 30개의 SSR 마커를 확인하였다. 개발된 175개 SSR 마커 중 20개 마커를 선발하여 'NC76'과 'GY14' 간 $F_2$ 분리 집단에 대한 연관지도를 작성하였으며 그 결과 13개의 마커가 예상했던 연관군에 일치하여 위치됨을 확인할 수 있었다. 따라서 본 연구에서 확인된 175개의 SSR 마커는 향 후 냉해 저항성 연관 마커 개발을 위한 오이 유전자 지도 작성 및 이를 통한 품종 육성에 크게 활용될 수 있을 것으로 기대된다.

• Animal cells and systems
• /
• 제1권1호
• /
• pp.123-128
• /
• 1997

The characteristics of allelic polymorphisms of the two (CA)n microsatellite (p599 and ㅅ599) markers spanning the long arm of chromosome 5 were studied in 52 DNA samples from unrelated inhabitants of Seoul (Korea) by using the polymerase chain reaction (PCR) to investigate differences in allele frequencies between Korean and Caucasian populations. The 6 alleles were observed for p599 (CA)n with a polymorphism informative content (PIC) value of 0.71 and 9 alleles for ㅅ599 (CA)n with a PIC value of 0.82. The observed heterozygote frequencies of the loci were estimated to 0.730 and 0.846, respectively. Several allele frequencies of two loci showed significant differences between Korean and Caucasian populations. Genotype data from the two loci were consistent with the Hardy-Weinberg equilibrium by x2 test. Linkage disequilibrium between p599 (CA)n and ㅅ599 (CA)n loci was observed in x2 test between the observed and expected frequency of allelic association. The probability of matching calculated at each locus was 0.104 for p599 (CA)n and 0.043 for ㅅ599 (CA)n, respectively. These results demonstrate the need to determine populationspecific allele frequency distributions for polymorphic markers when performing genetic linkage studies in racially defined several populations.

• 보존과학연구
• /
• 통권36호
• /
• pp.33-48
• /
• 2015

본 연구는 충청남도 아산시 탕정면 명암리 유적 9지점에서 발굴조사 된 고려 말에서 조선시대 초기 분묘 출토 인골 27개체를 대상으로 고유전학적 연구를 수행한 결과를 보고한 것이다. 실리카 추출 방법에 의하여 출토 인골로 부터 DNA를 추출한 후, 미토콘드리아 DNA(mtDNA)의 과변이지역을 9지역으로 나누어 PCR법에 의해 부분 증폭을 실시하였다. 증폭산물에 대한 염기서열 분석을 통하여 과변이지역의 변이형을 동정하였다. 그 결과 18개체의 mtDNA 분석이 가능하였으며, 아멜로제닌 유전자 분석에 의한 성결정은 M-26, M-29, M-37(L), M-49(R)의 4개체만이 분석되었고 모두 여성인 것으로 확인되었다.

• Journal of Microbiology and Biotechnology
• /
• 제30권11호
• /
• pp.1688-1696
• /
• 2020

Soil physical and chemical characteristics, soil potential denitrification rates (PDR), community composition and nirK-, nirS- and nosZ-encoding denitrifiers were studied by using MiSeq sequencing, quantitative polymerase chain reaction (qPCR), and terminal restriction fragment polymorphism (T-RFLP) technologies base on short-term (5-year) tillage field experiment. The experiment included four tillage treatments: conventional tillage with crop residue incorporation (CT), rotary tillage with crop residue incorporation (RT), no-tillage with crop residue retention (NT), and rotary tillage with crop residue removed as control (RTO). The results indicated that soil organic carbon, total nitrogen and NH₄⁺-N contents were increased with CT, RT and NT treatments. Compared with RTO treatment, the copies number of nirK, nirS and nosZ in paddy soil with CT, RT and NT treatments were significantly increased. The principal coordinate analysis indicated that tillage management and crop residue returning management were the most and the second important factors for the change of denitrifying bacteria community, respectively. Meanwhile, this study indicated that activity and community composition of denitrifiers with CT, RT and NT treatments were increased, compared with RTO treatment. This result showed that nirK, nirS and nosZ-type denitrifiers communities in crop residue applied soil had higher species diversity compared with crop residue removed soil, and denitrifying bacteria community composition were dominated by Gammaproteobacteria, Deltaproteobacteria, and Betaproteobacteria. Therefore, it is a beneficial practice to increase soil PDR level, abundance and community composition of nitrogen-functional soil microorganism by combined application of tillage with crop residue management.