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Distribution and Frequency of SSR Motifs in the Chrysanthemum SSR-enriched Library through 454 Pyrosequencing Technology (국화 SSR-enriched library에서 SSR 반복염기의 분포 및 빈도)

  • Moe, Kyaw Thu;Ra, Sang-Bog;Lee, Gi-An;Lee, Myung-Chul;Park, Ha-Seung;Kim, Dong-Chan;Lee, Cheol-Hwi;Choi, Hyun-Gu;Jeon, Nak-Beom;Choi, Byung-Jun;Jung, Ji-Youn;Lee, Kyu-Min;Park, Yong-Jin
    • Journal of the Korean Society of International Agriculture
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    • v.23 no.5
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    • pp.546-551
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    • 2011
  • Chrysanthemums, often called mums or chrysanths, belong to the genus Chrysanthemum, which includes about 30 species of perennial flowering plants in the family Asteraceae. We extracted DNA from Dendranthema grandiflorum ('Smileball') to construct a simple sequence repeat (SSR)-enriched library, using a modified biotin-streptavidin capture method. GS FLX (Genome Sequencer FLX System which provides the flexibility to perform the broad range of applications) sequencing (at the 1/8 run specification) resulted in 18.83 mega base pairs (Mbp) with an average read length of 280.06 bp. Sequence analyses of all SSR-containing clones revealed a predominance of di-nucleotide motifs (16,375, 61.5%) followed by tri-nucleotide motifs (6,616, 24.8%), tetra-nucleotide motifs (1,674, 6.3%), penta-nucleotide motifs (1,283, 4.8%), and hexa-nucleotide motifs (693, 2.6%). Among the di-nucleotide motifs, the AC/CA class was the most frequently identified (93.5% of all di-nucleotide types), followed by the GA/AG class (6.1%), the AT/TA class (0.4%), and the CG/GC class (0.03%). When we analyzed the distribution of different repeat motifs and their respective numbers of repeats, regardless of the motif class, of 100 SSR markers, we found a higher number of di-nucleotide motifs with 70 to 80 repeats; we also found two di-nucleotide motifs with 83 and 89 repeats, respectively, but their product lengths were within optimum size (297 and 300 bp). In future work, we will screen for polymorphisms of possible primer pairs. The results will provide a useful tool for assessing molecular diversity and investigating the population structure among and within Chrysanthemum species.

Overcoming taxonomic challenges in DNA barcoding for improvement of identification and preservation of clariid catfish species

  • Piangjai Chalermwong;Thitipong Panthum;Pish Wattanadilokcahtkun;Nattakan Ariyaraphong;Thanyapat Thong;Phanitada Srikampa;Worapong Singchat;Syed Farhan Ahmad;Kantika Noito;Ryan Rasoarahona;Artem Lisachov;Hina Ali;Ekaphan Kraichak;Narongrit Muangmai;Satid Chatchaiphan6;Kednapat Sriphairoj;Sittichai Hatachote;Aingorn Chaiyes;Chatchawan Jantasuriyarat;Visarut Chailertlit;Warong Suksavate;Jumaporn Sonongbua;Witsanu Srimai;Sunchai Payungporn;Kyudong Han;Agostinho Antunes;Prapansak Srisapoome;Akihiko Koga;Prateep Duengkae;Yoichi Matsuda;Uthairat Na-Nakorn;Kornsorn Srikulnath
    • Genomics & Informatics
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    • v.21 no.3
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    • pp.39.1-39.15
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    • 2023
  • DNA barcoding without assessing reliability and validity causes taxonomic errors of species identification, which is responsible for disruptions of their conservation and aquaculture industry. Although DNA barcoding facilitates molecular identification and phylogenetic analysis of species, its availability in clariid catfish lineage remains uncertain. In this study, DNA barcoding was developed and validated for clariid catfish. 2,970 barcode sequences from mitochondrial cytochrome c oxidase I (COI) and cytochrome b (Cytb) genes and D-loop sequences were analyzed for 37 clariid catfish species. The highest intraspecific nearest neighbor distances were 85.47%, 98.03%, and 89.10% for COI, Cytb, and D-loop sequences, respectively. This suggests that the Cytb gene is the most appropriate for identifying clariid catfish and can serve as a standard region for DNA barcoding. A positive barcoding gap between interspecific and intraspecific sequence divergence was observed in the Cytb dataset but not in the COI and D-loop datasets. Intraspecific variation was typically less than 4.4%, whereas interspecific variation was generally more than 66.9%. However, a species complex was detected in walking catfish and significant intraspecific sequence divergence was observed in North African catfish. These findings suggest the need to focus on developing a DNA barcoding system for classifying clariid catfish properly and to validate its efficacy for a wider range of clariid catfish. With an enriched database of multiple sequences from a target species and its genus, species identification can be more accurate and biodiversity assessment of the species can be facilitated.

Development of Marine Ecotoxicological Standard Methods for Ulva Sporulation Test (파래의 포자형성률을 이용한 해양생태독성시험 방법에 관한 연구)

  • Han, Tae-Jun;Han, Young-Seok;Park, Gyung-Soo;Lee, Seung-Min
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.13 no.2
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    • pp.121-128
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    • 2008
  • As an aquatic ecotoxicity test method, a bioassay using the inhibition of sporualtion of the green macroalga, Ulva pertusa, has been developed. Optimal test conditions determined for photon irradiance, pH, salinity and temperature were $100\;{\mu}mol{\cdot}m^{-2}{\cdot}s^{-1}$, $7{\sim}9$, $25{\sim}35\;psu$ and $15{\sim}20^{\circ}C$, respectively. The validity of the test endpoint was evaluated by assessing the toxicity of four metals (Cd, Cu, Pb, Zn) and elutriates of sewage or waste sludge collected from 9 different locations. When the metals were assayed, the $EC_{50}$ values indicated the following toxicity rankings: Cu ($0.062\;mg{\cdot}L^{-1}$) > Cd ($0.208\;mg{\cdot}L^{-1}$) > Pb ($0.718\;mg{\cdot}L^{-1}$) > Zn ($0.776\;mg{\cdot}L^{-1}$). When compared with other commonly used bioassays of metal pollution listed on US ECOTOX database, the sporualtion test proved to be the most sensitive. Ulva sporulation was significantly inhibited in all elutriates with the greatest and least effects observed in elutriates of sludge from industrial waste ($EC_{50}=6.78%$) and filtration bed ($EC_{50}=15.0%$), respectively. The results of the Spearman rank correlation analysis for $EC_{50}$ data versus the concentrations of toxicants in the sludge presented a significant correlation between toxicity and four heavy metals(Cd, Cu, Pb, Zn). The method described here is sensitive to toxicants, simple to use, easy to interpret and economical. It is also easy to procure samples and maintain cultures. The present method would therefore probably make a useful assessment of aquatic toxicity of a wide range of toxicants. In addition, the genus Ulva has a wide geographical distribution and species have similar reproductive processes, so the test method would have a potential application worldwide.

Colletotrichum Diversity within Different Species Complexes Associated with Fruit Anthracnose in South Korea and Their Fungicides In-Vitro Sensitivity (국내 과실 탄저병을 일으키는 종 복합체와 종 다양성 및 살균제 감수성)

  • Taehyun Chang;Oliul Hassan;Jong Yeob Jeon;Chi Hyun Kim;Dae Min Lee;Ju Sung Kim;Eun Chan Kang;Jaewon Kim
    • Research in Plant Disease
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    • v.29 no.4
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    • pp.345-362
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    • 2023
  • Anthracnose, caused by the Colletotrichum genus, comprises a significant number of plant pathogens and poses a major threat to fruit production worldwide, including South Korea. Colletotrichum species were identified associated with anthracnose in fruits such as apple, persimmon, plum, peach, jujube, walnut, and grape. A polyphasic approach, including morphology, multigene phylogenetics, and pathogenicity testing, was used. Additionally, the in-vitro sensitivity of identified Colletotrichum species to common fungicides was also evaluated. A total of nine Colletotrichum species within two complexes, namely gloeosporioides and acutatum, have been identified as the causal agents of anthracnose in common fruits in South Korea. In the gloeosporioides complex, we found Colletotrichumaenigma, C. fructicola, C. gloeosporioides, C. horii, C. siamense, and C. viniferum. Meanwhile, in the acutatum complex, C. fioriniae, C. nymphaeae, and C. orientalis were identified. Notably, C. fructicola, C. siamense, C. fioriniae, and C. nymphaeae were reported for the first time from apple, C. siamense, C. fioriniae and C. nymphaeae from plum, C. siamense, C. fructicola, and C. fioriniae frompeach, C. siamense and C. horii from persimmon, C. fioriniae from Omija (Schisandra), C. orientalis from walnut, C. nymphaeae from jujube, and C. aenigma, C. fructicola, and C. siamense fromgrape. Fungicide sensitivity tests revealed significant variation in the EC50 values among specific Colletotrichum species when exposed to different fungicides. Moreover, the same Colletotrichum species isolated from different host plants displayed varying sensitivity to the same fungicide.

In Vitro Anti-aging and Hair Follicle Dermal Papilla Cells Activation Effects of Usnea diffracta Vain Extract (송라 추출물의 세포 수준에서 항노화 및 모유두세포 활성화 효과)

  • Min Jeong Kim;Won Yeoung Choi;Hyun Woo Shim;Eun Jin Shin;Jung No Lee;Sung Min Park;Hwa Sun Ryu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.50 no.1
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    • pp.37-48
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    • 2024
  • Songla (Usnea diffracta Vain.) is one of the lichens belonging to the genus Usnea, and pharmacological activities such as antioxidant, antimicrobial, anti-inflammatory, anti-tumor and cardiovascular protection have been reported in previous studies, but its efficacy in skin and hair is not well known. In this study, the effect of Usnea diffracta extract (UDE) on anti-aging and dermal papilla cell proliferation was verified in vitro. As a result of the experiment, it was confirmed that the UDE significantly reduced the expression of MMP-1 and the activity of MAPKs (ERK, p38, JNK) and AP-1 (c-Fos, c-Jun), which were increased by UVA in HDFn. In addition, the UDE significantly increased the proliferation of HFDPC and significantly increased the mRNA expression of VEGF and KGF, which are hair growth factors. Accordingly, the phosphorylation of ERK/CREB involved in hair proliferation and expression of growth factors was increased in a concentration-dependent manner. The main component represented by the main peak was separated and purified using Prep LC by concentrating the UDE, which was confirmed as diffractaic acid through NMR and Mess analysis. Isolated diffractaic acid significantly reduced the expression of MMP-1 increased by UVA in HDFn and increased the proliferation of HFDPC in a concentration-dependent manner. The result suggest that UDE proved its usability as a natural cosmetic material with anti-aging and dermal papilla cell activation effects.

Trophic Level and Ecological Niche Assessment of Two Sympatric Freshwater Fish, Microphysogobio rapidus and Microphysogobio yaluensis Using Stable Isotope Analysis (안정동위원소 분석을 활용한 멸종위기종 여울마자와 동서종 돌마자의 영양단계 및 생태적 지위 평가)

  • Dae-Hee Lee;Hye-Ji Oh;Yerim Choi;Geun-Hyeok Hong;InHyuck Baek;Keun-Sik Kim;Kwang-Hyeon Chang;Ju-Duk Yoon
    • Korean Journal of Ecology and Environment
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    • v.57 no.1
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    • pp.39-50
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    • 2024
  • In ecosystems within limited resources, interspecific competition is inevitable, often leading to the competitive exclusion of inferior species. This study aims to provide foundational information for the conservation and restoration management of Microphysogobio rapidus by evaluating species' ecological response to biological factors within its habitat. To understand this relationship, we collected food web organisms from site where M. rapidus coexist with Microphysogobio yaluensis, a specie ecologically similar to M. rapidus, and evaluated the trophic levels (TL), isotopic niche space (INS), and the overlap of INS among fishes within the habitat using stable isotope analysis. Our analysis revealed that the M. rapidus exhibited a higher TL than M. yaluensis, with TL of 2.6 and 2.4, respectively. M. yaluensis exhibited a broad INS, significantly influencing the feeding characteristics of most fish. Conversely, M. rapidus showed a narrow INS and asymmetric feeding relationships with other species, in habitats with high competition levels. This feeding characteristics of M. rapidus indicate that the increase in competitors sharing the similar resources lead to a decrease in available resources and, consequently, is expected to result in a decrease in their density.

Changes in Fermentation Characteristics and Bacterial Communities of Whole Crop Rice Silage during Ensiling Period (저장기간에 따른 사료용 벼 사일리지의 발효특성 및 미생물상 변화)

  • Mirae Oh;Hyung Soo Park;Bo Ram Choi;Jae Hoon Woo;Seung Min Jeong;Ji Hye Kim;Bae Hun Lee
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.44 no.1
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    • pp.1-5
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    • 2024
  • Understanding changes in fermentation characteristics and microbial populations of forage silage during ensiling is of interest for improving the nutrient value of the feed for ruminants. This study was conducted to investigate the changes in fermentation characteristics and bacterial communities of whole crop rice (WCR) silage during the ensiling period. The chemical compositions, pH, organic acids and bacterial communities were evaluated at 0, 3, 6, and 12 months after ensiling. The bacterial communities were classified at both the genus and species levels. The dry matter content of WCR silage decreased with the length of storage (p<0.05), but there was no significant difference in crude protein and NDF contents. Following fermentation, the pH level of WCR silage was lower than the initial level. The lactic acid content remained at high levels for 3 to 6 months after ensiling, followed by a sharp decline at 12 months (p<0.05). Before fermentation, the WCR was dominated by Weissella (30.8%) and Pantoea (20.2%). Growth of Lactiplantibacillus plantarum (31.4%) was observed at 3 months after ensiling. At 6 months, there was a decrease in Lactiplantibacillus plantarum (10.2%) and an increase in Levilactobacillus brevis (12.8%), resulting in increased bacteria diversity until that period. The WCR silage was dominated by Lentilactobacillus buchneri (71.2%) and Lacticaseibacillus casei (27.0%) with a sharp reduction in diversity at 12 months. Overall, the WCR silage maintained satisfactory fermentation quality over a 12-month ensiling period. Furthermore, the fermentation characteristics of silage were found to be correlated to bacterial microbiome.

Identifying Distribution Areas and Population Sizes for the Conservation of the Endangered Species Odontobutis obscura (멸종위기종 남방동사리의 보전을 위한 상세 분포 지역 및 개체군 크기 파악)

  • Jeong-Hui Kim;Sang-Hyeon Park;Seung-Ho Baek;Chung-Yeol Baek
    • Korean Journal of Ecology and Environment
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    • v.57 no.2
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    • pp.102-110
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    • 2024
  • This study presents a fine scale distribution of the endangered species, Odontobutis obscura, through field surveys and literature reviews. Using the mark-recapture method, the population size in major habitats was determined. Field surveys conducted on 18 streams in Geoje Island revealed that the O. obscura was only found in the main streams and tributaries of the Sanyang, Gucheon, and Buchun Streams, which are part of the Sanyang Stream watershed. The O. obscura exhibited relative abundances ranging from 0.5% to 35.3% at different locations, with certain spots showing higher relative abundances (18.8% to 35.3%), indicating major habitat areas. A review of six literature studies confirmed the presence of the O. obscura, although there were differences in occurrence status depending on the purpose, scope, and duration of the studies. Combining the results of field and literature surveys, it was found that the O. obscura inhabits the main and tributary streams of the Sanyang, Gucheon, and Buchun Streams, from the upper to lower reaches. Currently, the O. obscura population in the Sanyang Stream watershed maintains a stable habitat, but its limited distribution range suggests potential issues such as genetic diversity deficiency in the long term. The population size of the O. obscura was confirmed at two specific locations, with densities of 0.5 to 1.5 individuals per m2. The average movement distance from the release point was 13.1 m, indicating the limited mobility characteristic of ambush predators. Understanding the distribution and population size of endangered species is the first step towards their conservation and protection. Based on this information, further research could significantly contribute to the conservation of the O. obscura.

Evaluation of anti-inflammatory efficacy of Lacticaseibacillus rhamnosus L22-FR28 (KACC 92513P) isolated from infant feces and its Oenanthe javanica ferments (영아분변 유래 Lacticaseibacillus rhamnosus L22-FR28(KACC 92513P) 균주와 미나리 발효물의 항염증 효능 평가)

  • Seoyeon Kwak;Hee-Min Gwon;Soo-Hwan Yeo;So-Young Kim
    • Food Science and Preservation
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    • v.31 no.3
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    • pp.474-485
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    • 2024
  • The purposes of this study were to isolate the potential Lacticaseibacillus spp. from the feces of infants before weaning, to investigate the safety of antibiotics resistance and beta-haemolysis, and to evaluate the anti-bacterial and anti-inflammatory effects between the selected strains and Oenanthe javanica (Oj) fermented by them. As a result of analyzing the intestinal microbial community among the stools of four infants, the genus Bifidobacterium was the most dominant, but Lacticaseibacillus (L.) rhamnosus was the most frequently isolated because of the easy culture. Nine test strains, including Lactobacillus rhamnosus LGG (ATCC 53103) as the positive control, were sensitive against 8 kinds of antibiotics without vancomycin in comparison with the cut-off values at the European Food Safety Authority (EFSA), and there was no hemolysis. In the antibacterial activity experiment, the Lacticaseibacillus rhamnosus L22-FR28 (L28, KACC 92513P) strain and Oj+L28 ferment showed significantly (p<0.05) higher activities than LGG against Bacillus cereus and Staphylococcus aureus. Additionally, these decreased the activity of the NF-kB/AP-1 transcription factor and inhibited the nitric oxide and cytokines (TNF-α and IL-6) produced in macrophage RAW cells stimulated by lipopolysaccharide (LPS). Consequently, the L. rhamnosus L28 strain and Oenanthe javanica+L. rhamnosus L28 (Oj+L28) ferment selected with the high anti-inflammatory effect will improve health functionality after more research, such as the verification of animal level and identification of mechanism on an anti-inflammatory.

Characteristics of New Microsporidia S80 Isolated from Silkworm, Bombyx mori L. in Korea (가잠(家蠶)으로부터 분리(分離)된 새로운 Microsporidia S80의 특성(特性))

  • Lim, Jong Sung;Cho, Sae Yun
    • Current Research on Agriculture and Life Sciences
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    • v.1
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    • pp.67-83
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    • 1983
  • The new microsporidia S80 isolated from, Bombyx mori L. in Korea showed ovoid in the morphology of the spores and the size were measured $2.9{\pm}0.28{\mu}$ in length and $1.7{\pm}0.29{\mu}$ width. No other microsporidian spore like this has not been so far isolated from Silkworm. The length of the polar filament extruded in hydrogen peroxide ($H_2O_2$) at $30^{\circ}C$ was $26{\mu}$ of a round cytoplasm on the top. The spores were partly stained with Giemsa, Safranin-O and Gram as the same staining properties as Nosema bombycis, Microsporidia K 79 and other microsporidian spores. The fine structures were observed under scanning eleceron microscope through ultrathin sectioning. The spore wall was composed of three layers ; the thin exospore of an electron dense rippled layer, the thick electron lucent endospore which was thinning considerably at the polar filament insertion point, and the inner limiting membrane. Polar cap present at the sporeapex, with a long polar filament of 12-13 coils, subtending angle of $60^{\circ}$ to spore axis, which is tubular made up of a multilayered and are a benes core, light ring structure enclosing the dance core, the dark ring structure enclosing the inner light ring structure and the other than and light ring structure bounded from cytoplasm. Lamellate polaroplast occupied the anterior part of the spore, and the two neclei with dense nucleoplasm bounded by a double nuclear envelope were cited in the slight downer middle portion of spore. From the characteristics of the shape, size and fine structures, it is certain to reason the Microsporidia S80 belong to the phylum Microspora, class Microspora, order Microsporida, order Microsporida. The shape of two nuclei cited seems to be genus Nosema, but in the classification for the suborder it should be defined wheather pansporoblasts be formed or not and for the genis especial attempts have been made to define the characters which distinguish the disporous genera in the life cycle. Survey through the infection of the bad cocoons during 1980 to 1982 in South Korea the areas contaminated with new microsporidia were revealed 5 provinces of Kyung-Gi, Kang-Won, Chung-Nam and Chun-Nam. Pathological effects inoculated per os at second instar larvae of silkworm, the LD 50 was $7.1{\times}10^7/ml$ as lower pathogenecity than that of Nosema bombycis Naegeli of $1.2{\times}10_7/ml$. While on the other hand the inoculation of the microsporidia at fourth instar larvae lowerd the whole cocoon weight and cocoon shell weight and significant at 1% level. The microsporidia S80 defined it can not be transmitted transovarially from the result of predictive and collective examination of 21 egg batches from the infected female moth.

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