• Title/Summary/Keyword: Internal transcribed spacer rDNA

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Molecular Identification of a Sea Anemone (Cnidaria: Anthozoa: Actiniaria) Obtained in Gijang, Busan (부산 기장에서 채집된 말미잘의 분자생물학적 방법을 이용한 동정)

  • Yoo, Sang Joon;Kim, Do-Hyung
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.50 no.4
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    • pp.447-452
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    • 2017
  • In this study, we tried to identify a sea anemone collected from the coast of Gijang, Busan. The anemone was morphologically similar to species belonging to the genus Anthopleura, but its morphological characteristics did not allow for confirmed identification to species level. Multiple genes from mitochondrial cytochrome oxidase III, 12S and 16S rRNA, and nuclear 18S and 28S rRNA, were amplified for multilocus sequence typing (MLST) analysis using genomic DNA extracted from the sampled anemone and a different primer set. Based on the MLST analysis, the anemone obtained in this study was identified as Anthopleura artemisia. Also, the sequence of internal transcribed spacer-2 was most closely related to A. artemisia, indicating that this single region might be useful for anemone identification. This study shows significance of molecular identification for sea anemones, and will be helpful in studies of sea anemone identification using genotyping-by-sequencing.

Molecular Identification and Technological Properties of Acetic Acid Bacteria Isolated from Malatya Apricot and Home-Made Fruit Vinegars

  • Buyukduman, Eda;Kirtil, Hatice Ebrar;Metin, Banu
    • Microbiology and Biotechnology Letters
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    • v.50 no.1
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    • pp.81-88
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    • 2022
  • Acetic acid bacteria (AAB) are versatile organisms involved in the production of variety of fermented foods, such as vinegar and kombucha, and products of biotechnological relevance, such as bacterial cellulose. In the present study, Malatya apricot, a variety with protected designation of origin (PDO), and vinegar samples produced using various fruits were used to isolate AAB. The 19 AAB isolates obtained were typed using (GTG)5 fingerprinting, and the ones selected were identified by sequencing either 16S rDNA alone or in combination with 16S-23S rRNA internal transcribed spacer region or ligA gene. While all apricot isolates (n = 10) were Gluconobacter cerinus, vinegar isolates (n = 9) were composed of Komagataeibacter saccharivorans, Acetobacter syzygii, and possible two new species of AAB, Komagataeibacter sp., and Gluconobacter sp. (GTG)5 fingerprinting showed the presence of several genotypes of G. cerinus in the apricot samples. Screening for some technologically relevant properties, including thermotolerance, ethanol tolerance, and cellulose production capability, showed that all Komagataeibacter and some Gluconobacter isolates could tolerate the temperature of 35℃, and that vinegar isolates could tolerate up to 8% ethanol. One isolate, Komagataeibacter sp. GUS3 produced bacterial cellulose (1 g/l) and has the potential to be used for cellulose production.

Quantification of triterpenes in Centella asiatica cultivated in a smart farm, and their effect on keratinocyte activation (스마트팜 재배 병풀의 triterpenes 정량 및 각질형성세포 활성화 효과)

  • Jin Hong Park;Seong Min Jo;Da Hee Lee;Youngmin Park;Hwan Bong Chang;Tae Jin Kang;Kiman Lee
    • Food Science and Preservation
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    • v.30 no.3
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    • pp.483-491
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    • 2023
  • This study aimed to compare the bioactive compounds in Centella asiatica (C. asiatica) cultivated in a smart farm and a field and their effects on human keratinocyte cells. C. asiatica was collected in Jeju-do, Korea, and cultured in a smart farm and a field. The main bioactive compounds in the two differentially cultured C. asiatica were identified, and their activation in keratinocytes were assessed. Amplification and sequencing of the internal transcribed spacer (ITS) DNA in the nucleus and psbA-H DNA in the chloroplast were performed for species analysis. A comparison of DNA of plants reported in the NCBI GenBank was performed. The ITS DNA and psbA-H DNA sequences of C. asiatica cultivated in a smart farm and a field were consistent with No. MH768338.1 and No. JQ425422.1, respectively. Analysis of the triterpenes was performed using high performance liquid chromatography (HPLC) and as a result, C. asiatica cultured in a smart farm had more triterpenes than those cultured in a field. The effects of C. asiatica grown in a smart farm on cell proliferation and scratch recovery in HaCaT cells were greater than those grown in a field. These results suggest that C. asiatica cultivated in a smart farm can be effectively utilized as a health functional food.

ITS sequence variations in common ragweed and giant ragweed (돼지풀 및 단풍잎돼지풀의 ITS 염기서열 변이)

  • Kim, Young-Dong;Park, Chong-Wook;Sun, Byung-Yun;Kim, Ki-Joong;Lee, Eun-Ju;Kim, Sung-Hee
    • Korean Journal of Plant Taxonomy
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    • v.35 no.4
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    • pp.273-285
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    • 2005
  • Genetic variation of notorious invasive plants, common ragweed (Ambrosia artemisiifolia L.) and giant ragweed (Ambrosia trifida L.) were examined using the internal transcribed spacer (ITS) sequences of nuclear ribosomal DNA. A total of 18 different ITS types were detected from 156 individuals of common ragweed sampled mainly from the southern part of Korean peninsula whereas four types were identified from 46 individuals of giant ragweed. High sequence diversity observed from common ragweed in Korean populations was interpreted as multiple introduction. Genetic recombination was suggested as possible method for the production of some of the ITS types while point mutation was mainly responsible for the origin of the sequence diversity. This study provided some of basic genetic information needed for understanding of the evolutionary process in ragweed during invasion.

Isolation of Fungal Deteriogens Inducing Aesthetical Problems and Antifungal Calcite Forming Bacteria from the Tunnel and Their Characteristics (터널에서 미학적 문제를 야기하는 진균 및 항진균 활성을 가진 탄산칼슘 형성세균의 분리와 특성)

  • Park, Jong-Myong;Park, Sung-Jin;Ghim, Sa-Youl
    • Microbiology and Biotechnology Letters
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    • v.39 no.3
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    • pp.287-293
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    • 2011
  • The purpose of this study was to isolate and characterize fungal deteriogens, which induce discoloration of the cement tunnel, and calcite forming bacteria (CFBs), which have antifungal activity against fungal deteriogens. Isolation of mold, bacteria and yeast was performed using several solid media and partially identified using internal transcribed spacer (ITS); 5.8S rRNA gene sequencing and 16s rDNA sequencing. A total of 19 microbial strains were identified with the most widely distributed fungal strain being Cladospirum sphaerospermum. In addition, five bacteria derived from the tunnel were identified as CFBs. Amongst the latter, Bacillus aryabhatti KNUC205 exhibited antifungal activity against Cladospirum sphaerospermum KNUC253 and Aspergillus niger KCTC6906 as concentrated filtered supernatants.

Genetic Diversity of Didymella bryoniae for RAPD Profiles Substantiated by SCAR Marker in Korea

  • Shim, Chang-Ki;Seo, Il-Kyo;Jee, Hyeong-Jin;Kim, Hee-Kyu
    • The Plant Pathology Journal
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    • v.22 no.1
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    • pp.36-45
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    • 2006
  • Twenty isolates of Didymella bryoniae were isolated from infected cucurbit plants in various growing areas of southern Korea in 2001 and 2002. Random Amplified Polymorphic DNA (RAPD) group [RG] I of D. bryoniae was more virulent than RG IV to watermelon. Virulence of the RG I isolate was strong to moderate to cucumber, whereas that of the RG IV varied from strong, moderate to weak. Two hundred seventy-three amplified fragments were produced with 40 primers, and were analyzed by a cluster analysis using UPGMA method with an arithmetic average program of NTSYSPC. At the distance level of 0.7, two major genomic DNA RAPD groups were differentiated among 20 isolates. The RG I included 7 isolates from watermelon and one isolate from melon, whereas the RG IV included 12 isolates from squash, cucumber, watermelon and melon. Amplification of internal transcribed spacer (ITS) region and small subunit rRNA region from the 20 isolates yielded respectively a single fragment. Restriction pattern with 12 restriction enzymes was identical for all isolates tested, suggesting that variation in the ITS and small subunit within the D. bryoniae were low. Amplification of the genomic DNAs of the tested isolates with the sequence characterized amplified regions (SCAR) primer RG IF-RG IR specific for RG I group resulted in a single band of 650bp fragment for 8 isolates out of the 20 isolates. Therefore, these 8 isolates could be assigned into RG I. The same experiments done with RG IIF-RG IIR resulted in no amplified PCR product for the 20 isolates tested. An about 1.4 kb-fragment amplified from the RG IV isolates was specifically hybridized with PCR fragments amplified from genomic DNAs of the RG IV isolates only, suggesting that this PCR product could be used for discriminating the RG IV isolates from the RG I isolates as well other fungal species.

The Incidence of Alternaria Species Associated with Infected Sesamum indicum L. Seeds from Fields of the Punjab, Pakistan

  • Nayyar, Brian Gagosh;Woodward, Steve;Mur, Luis A.J.;Akram, Abida;Arshad, Muhammad;Naqvi, S.M. Saqlan;Akhund, Shaista
    • The Plant Pathology Journal
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    • v.33 no.6
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    • pp.543-553
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    • 2017
  • Sesame (Sesamum indicum) is an important oil seed crop of Asia. Yields can be negatively impacted by various factors, including disease, particularly those caused by fungi which create problems in both production and storage. Foliar diseases of sesame such as Alternaria leaf blight may cause significant yield losses, with reductions in plant health and seed quality. The work reported here determined the incidence of Alternaria species infecting sesame seeds grown in the Punjab, Pakistan. A total of 428 Alternaria isolates were obtained from 105 seed samples and grouped into 36 distinct taxonomic groups based on growth pattern and morphological characters. Isolation frequency and relative density of surface sterilized and non-surface sterilized seeds showed that three isolates (A13, A47 and A215) were the most common morphological groups present. These isolates were further identified using sequencing of the Internal Transcribed Spacer (ITS) region of ribosomal DNA (rDNA) and the Alternaria major allergen gene (Alt a 1). Whilst ITS of rDNA did not resolve the isolates into Alternaria species, the Alt a 1 sequences exhibited > 99% homology with Alternaria alternata (KP123850.1) in GenBank accessions. The pathogenicity and virulence of these isolates of Alternaria alternata was confirmed in inoculations of sesame plants resulting in typical symptoms of leaf blight disease. This work confirms the identity of a major source of sesame leaf blight in Pakistan which will aid in formulating effective disease management strategies.

Morphological and genetic characterization and the nationwide distribution of the phototrophic dinoflagellate Scrippsiella lachrymosa in the Korean waters

  • Lee, Sung Yeon;Jeong, Hae Jin;You, Ji Hyun;Kim, So Jin
    • ALGAE
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    • v.33 no.1
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    • pp.21-35
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    • 2018
  • The phototrophic dinoflagellate genus Scrippsiella is known to have a worldwide distribution. Here, we report for the first time, the occurrence of Scrippsiella lachrymosa in Korean waters. Unlike the other stains of S. lachrymosa whose cultures had been established from cysts in the sediments, the clonal culture of the Korean strain of S. lachrymosa was established from motile cells. When the sulcal plates of S. lachrymosa, which have not been fully described to date, were carefully examined using scanning electron microscopy, the Korean strain of S. lachrymosa clearly exhibited the anterior sulcal plate (s.a.), right sulcal plate (s.d.), left sulcal plate (s.s.), median sulcal plate (s.m.), and posterior sulcal plate (s.p.). When properly aligned, the large subunit (LSU) rDNA sequence of the Korean strain of S. lachrymosa was ca. 1% different from those of two Norwegian strains of S. lachrymosa, the only strains for which LSU sequences have been reported. The internal transcribed spacer (ITS) rDNA sequence of the Korean strain of S. lachrymosa was also ca. 1% different from those of the Scottish and Chinese strains and 3% different from those of the Canadian, German, Greek, and Portuguese strains. Thus, the Korean S. lachrymosa strain has unique LSU and ITS sequences. The abundances of S. lachrymosa in the waters of 28 stations, located in the East, West, and South Sea of Korea, were quantified in four seasons from January 2016 to October 2017, using quantitative real-time polymerase chain reaction method and newly designed specific primer-probe sets. Its abundances were >$0.1cells\;mL^{-1}$ at eight stations in January and March 2016 and March 2017, and its highest abundance in Korean waters was $26cells\;mL^{-1}$. Thus, S. lachrymosa has a nationwide distribution in Korean waters as motile cells.

Genetic Diversity and Pathogenicity of Cylindrocarpon destructans Isolates Obtained from Korean Panax ginseng

  • Song, Jeong Young;Seo, Mun Won;Kim, Sun Ick;Nam, Myeong Hyeon;Lim, Hyoun Sub;Kim, Hong Gi
    • Mycobiology
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    • v.42 no.2
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    • pp.174-180
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    • 2014
  • We analyzed the genetic diversity of Cylindrocarpon destructans isolates obtained from Korean ginseng (i.e., Panax ginseng) roots by performing virulence tests and nuclear ribosomal gene internal transcribed spacer (ITS) and mitochondrial small subunit (mt SSU) rDNA sequence analysis. The phylogenetic relationship analysis performed using ITS DNA sequences and isolates from other hosts helped confirm that all the Korean C. destructans isolates belonged to Nectria/Neonectria radicicola complex. The results of in vivo and ex vivo virulence tests showed that the C. destructans isolates could be divided into two groups according to their distinctive difference in virulence and the genetic diversity. The highly virulent Korean isolates in pathogenicity group II (PG II), together with foreign isolates from P. ginseng and P. quinquefolius, formed a single group. The weakly virulent isolates in pathogenicity group I, together with the foreign isolates from other host plants, formed another group and exhibited a greater genetic diversity than the isolates of PG II, as confirmed by the mt SSU rDNA sequence analysis. In addition, as the weakly virulent Korean isolates were genetically very similar to the foreign isolates from other hosts, they were likely to originate from hosts other than the ginseng plants.

Isolation and Identification of Wild Yeasts from Freshwaters and Soils of Nakdong and Yeongsan River, Korea, with Characterization of Two Unrecorded Yeasts (낙동강과 영산강 담수와 주변 토양으로부터 야생효모의 분리 및 동정)

  • Han, Sang-Min;Kim, Ha-Kun;Lee, Hyang-Burm;Lee, Jong-Soo
    • The Korean Journal of Mycology
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    • v.44 no.4
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    • pp.350-354
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    • 2016
  • Diverse wild yeast were isolated from freshwaters and soils of Nakdong and Yeongsan rivers in Korea and identified by the comparison of polymerase chain reaction-amplified nucleotide sequences of the internal transcribed spacer region (including the 5.8S rRNA) and D1/D2 regions of 26S rDNA, using BLAST. In total, 15 strains belonging to 9 species were isolated from 25 samples, out of which Aureobasidium pullulans and Cryptococcus bestiolae were dominant. Candida ghanaensis JSF0127 and Meira geulakonigii JSF0130 were identified as unrecorded yeasts, for which their mycological characteristics were investigated. These unrecorded yeasts formed ascospores and grew in yeast extract peptone dextrose medium containing 5% NaCl.