• Korean Journal of Microbiology
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• v.33 no.2
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• pp.149-156
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• 1997

We have employed comparative RFL,P(Restriction Fragment Ixngth Pol~iniorphism) analysis and molecular phylogenetic techniques to investigate the diversity of uncultured microorganisms associated with the anaerobic PCP degradation in PCP-adapted enrichment cultures inoculated by samples from anaerobic cewage sludgc(Jangrim, Pusan) and leachate of landfill site(Kimhae). 16s rDNA cloncs were obtairted by PCR amplification of mixed population DNAs extracted directly from the nonactive and active stage ol each PCP-adapted culture. After three rounds of comparative RFLP analyses. two RFLP types. designated as Ala and Hld, were found prevalent and common in both active stage samples. Thc analysis of phylogenctic diversity bawd on the 5'-terminal 180 nt of sequences from whole clones of the Ala and Bld RFLP types showed close similarity among themselves. In case of Bld clones, 7XQ of them shared identical sequences. Thcse resuliq suggest that the clones of both RFLP types wcre originated from highly affiliated microorganisms which are e~iriched as a result of metabolic activity to PCP. The full-length 16s rRNA sequence of each representative clone from both RFLP types was determined. and an Ala clone w i n found to he related to Clo.strrdiurn ulfutzac~(Genk~ank No. Z69203) and a Bld clone to Thermobacteroides proteolyticus(Genbank No. X09335), with sequence similarities of 89%' and 97%. respectively.

• Journal of Genetic Medicine
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• v.2 no.2
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• pp.53-57
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• 1998

Spinal muscular atrophy (SMA) type I is a common severe autosomal recessive inherited neuromuscular disorder that has been mapped to chromosome 5q11.2-13.3. The survival motor neuron (SMN) gene, a candidate gene, is known to be deleted in 96% of patients with SMA type I. Presently, PCR and single strand conformation polymorphism (PCR-SSCP) analyses have been made possible for application to both archival slides and paraffin-embedded tissues. Archival materials represent valuable DNA resources for genetic diagnosis. We applied these methods for the identification of SMN gene of SMA type I in archival specimens for the prenatal diagnosis. In this study, we performed the prenatal diagnosis with chorionic villus sampling (CVS) cells on two women who had experienced neonatal death of SMA type I. DNA extraction was done from archival slide and tissue materials and PEP-PCR was performed using CVS cells. In order to identify common deletion region of SMN and neuronal apoptosis-inhibitory protein (NAIP) genes, cold PCR-SSCP and PCR-restriction site assay were carried out. Case 1 had deletions of the exons 7 and 8, and case 2 had exon 7 only on the telomeric SMN gene. Both cases were found to be normal on NAIP gene. These results were the same for both CVS and archival biopsied specimens. In both cases, the fetuses were, therefore, predicted to be at very high risk of being affected and the pregnancy were terminated. These data clearly demonstrate that archival slide and paraffin-embedded tissues can be a valuable source of DNA when the prenatal genetic diagnosis is needed in case any source for genetic analysis is not readily available due to previous death of the fetus or neonate.

• Horticultural Science & Technology
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• v.34 no.6
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• pp.912-923
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• 2016

Watermelon production is often limited by powdery mildew in areas with a large daily temperature range. Development of resistant watermelon cultivars can protect against powdery mildew; however, little is known about the characteristics of its causal agents. Here, we identified the genus and race of a causal pathogen of powdery mildew in Ansung province of South Korea, and developed molecular markers for the generation of resistant watermelon cultivars. The causal pathogen was determined to be Podosphaera xanthii based on multiple sequence alignments of internal transcribed spacers (ITS) of rDNA. The physiological race was identified as 1W, and the Ansung isolate was named P. xanthii 1W-AN. Following inoculation with the identified P. xanthii 1W-AN, we found inheritance of the resistant gene fitting a single dominant Mendelian model in a segregated population ('SBA' ${\times}$ PI 254744). To develop molecular markers linked to fungus-resistant loci, random amplified polymorphic DNA (RAPD) was accomplished between DNA pooled from eight near-isogenic lines (NILs; $BC_4F_6$), originated from PI 254744 and susceptible 'SBB' watermelon. After sequencing bands from RAPD were identified in all eight NILs and PI254744, 42 sequence-characterized amplifiedregion (SCAR) markers were developed. Overall, 107 $F_2$ plants derived from $BC_4F_6$ NIL-1 ${\times}$ 'SBB' were tested, and one SCAR marker was selected. Sequence comparison between the SCAR marker and the reference watermelon genome identified three Nco I restriction enzyme sites harboring a single nucleotide polymorphism, and codominant cleavage-amplified polymorphic site markers were subsequently developed. A CAPS marker was converted to a high-resolution melt (HRM) marker, which can discriminate C/T SNP (254PMR-HRM3). The 254PMR-HRM3 marker was evaluated in 138 $F_{2:3}$ plants of a segregating population ('SBA' ${\times}$ PI254744) and was presumed to be 4.3 cM from the resistance locus. These results could ensure P. xanthii 1W-AN resistance in watermelon germplasm and aid watermelon cultivar development in marker-assist breeding programs.

• Korean Journal of Environment and Ecology
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• v.33 no.2
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• pp.202-215
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• 2019

This study was conducted to investigate the spawning preference of the Acheilognathinae fishes in relation to the shell size of host mussels after identifying the species of eggs and fries in the host mussel using our recently developed RFLP (Restriction Fragment Length Polymorphism) molecular marker at four sites [Hongcheon Naechoncheon (HN) and Deokchicheon (HD) from the North Han River basin and Jeongseon Goljicheon (JG) and Joyanggang (JJ) from the South Han River] in South Korea during May in each year between 2015 and 2018. The Acheilognathinae fish observed in the studied sites included one species (Acheilognathus signifer) in HN and JG, three species (Rhodeus uyekii, A. signifer, and Acheilognathus yamatsutae) in HD, and two species (A. signifer and Acheilognathus yamatsutae) in JJ, and we collected 982 host mussels (Unio douglasiae sinuolatus) that inhabited in all four sites. Using the RFLP molecular marker, we confirmed 46 eggs and fry of the Acheilognathinae fish (454 A. signifer, 43 Acheilognathus yamatsutae, and 149 Acheilognathus yamatsutae) in Unio douglasiae sinuolatus (N=163; 16.6%). We compare the average shell length, shell height, and shell width of mussels with [presence] eggs/fry and mussels without [absence] eggs/fry to examine the spawning preference according to the size of host mussels in each site. The results show that the shell length (1.98 mm), shell height (0.85 mm), and shell width (0.73 mm) of mussels with the eggs/fry were significantly larger (Mann-Whitney U test, P=0.002; difference=1.98 mm) than those of mussel without eggs/fry in HD where three species cohabitated. Although the shell length, shell height, and shell width of mussels with the eggs/fry were larger also in the other three sites, the difference was not statistically significant. In addition, we analyzed the mean number of spawned eggs and fry of each species and found $9.31{\pm}5.94$ R. uyekii, $2.86{\pm}2.45$ A.signifer, and $2.50{\pm}1.32$ A. yamatsutae. R. uyekii spawned 6.45-6.81 more eggs than A.signifer and A. yamatsutae on average per mussel, and it was statistically significant (Kruskal-Wallis test, P < 0.001). These findings indicate that the three species of Acheilognathinae fish tend to prefer larger mussels as their spawning hosts, and this tendency increases when the number of cohabitating bitterling fish species increases. Moreover, A.signifer and A. yamatsutae spawned a smaller number of eggs evenly in more host mussels while R. uyekii spawned many eggs on relatively fewer mussels. We found mussels (N=4) having the eggs/fry of two coexisting species, A. signifier and A. yamatsutae in HD and JJ where more than two bitterling fish species occurred. It suggests the interspecific competition taking place between the Acheilognathinae fishes for utilizing the same resource of mussels for spawning when two or more species cohabitate. This study is expected help to understand better the spawning patterns and reproductive ecology of the Acheilognathinae fishes, which will provide insightful information for advancing our understanding of their ecological relationships - mutualism or host-parasitism - with host mussels.