• Title/Summary/Keyword: 분자 계통분석

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Molecular Identification of Pseudanabaena Strains and Analysis of 2-MIB Production Potential in the North Han River System (북한강 수역에 분포하는 Pseudanabaena 균주의 동정 및 2-MIB 생산 잠재성 분석)

  • Kim, Keonhee;Lee, Sejin;Seo, Kyunghwa;Hwang, Soon-Jin
    • Korean Journal of Ecology and Environment
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    • v.53 no.4
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    • pp.344-354
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    • 2020
  • Identification of the target species of 2-MIB (2-methyllisoborneol) production is crucial in the management of off-flavor problem in the freshwater system. This study was conducted to identify 2-MIB-producing Pseudanabaena strains occurring in the North Han River system using molecular genetic method. Eleven phenotypes of Pseudanabaena were isolated from several mainstream sites of the North Han River, including Sambong-ri, Joam-myun, and Lake Uiam areas. Despite of morphological similarity of the strains, the phylogenetic analysis using 16S rDNA classified them into different species with low genetic similarity (40~55%). Isolated Pseudanabaena strains were converged to four species; Pseudanabaena cinerea, P. yagii, P. mucicola, and P. redekei. Among them, the 2-MIB synthesizing gene (mibC) was detected in P. cinerea, P. yagii, and P. redekei. However, actual 2-MIB production was detected only in P. cinerea and P. redekei based on gas chromatography analysis. This study is the first report of the molecular identification of Pseudanabaena strains and their 2-MIB production potential in Korea. The results of this study provides an evidence of species diversity of Pseudanabaena occurring in the North Han River.

Molecular Phylogeny of the Genera Staurastrum and Staurodesmus (Zygnematophyceae, Streptophyta) Based on Nuclear (18S rDNA) and Chloroplast Gene (atpB) Sequences (핵(18S rDNA)과 엽록체 유전자(atpB) 분석을 통한 Staurastrum속과 Staurodesmus속 (Zygnematophyceae, Streptophyta)의 분자 계통학적 연구)

  • Moon, Byeong-Ryeol;Lee, Ok-Min
    • ALGAE
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    • v.22 no.1
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    • pp.1-10
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    • 2007
  • To gain insights into the phylogenetic relationships of genus Staurastrum and Staurodesmus, we analyzed nuclearencoded small subunit rDNA of 82 strains, and chloroplast atpB gene sequences of 44 strains belonging to three genera (Staurastrum, Staurodesmus, Cosmarium). Excluding the Staurastrum muticum and S. orbiculare, forty five strains of genus Staurastrum formed a well supported clade. It was shown that with no cell wall sculpture and processes, these two species have a strong phylogenetic relationship with genus Staurodesmus. Therefore, it is strongly recommended to transfer Staurastrum without processes and cell wall sculpture into Staurodesmus. S. obsoletus is a taxa that is transferred from Cosmarium. But, from this study, it has shown a phylogenetic relationship with Cosmarium. Therefore, this species is strongly recommended to transfer back to Cosmarium instead of Staurodesmus. As it was studied before, genus Staurastrum has shown monophyletic. Since the genus taurodesmus groups with Cosmarium, they were shown to be polyphyletic.

Nucleotide Analysis of 185 rRNA and Molecular Phylogeny of the Korean Decapods (하국산 십각류의 18S 리보솜 RNA의 염기분석과 분자계통에 관한 연구)

  • Kim, Won
    • The Korean Journal of Zoology
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    • v.35 no.1
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    • pp.80-86
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    • 1992
  • The nucleotide sequences of 185 rRNAs of the five Korean decapods were partially determined by the direct sequencing method using the reverse transcriptase. ne average GC content of five species was 51.1% which is higher than that of yeast(45.0%) and lower than those of frog (53.0%) and rat (55.6%). This result follows the general patterns of the GC content in the nucleotides of the nucleic acid shown among the various phylogenetic groups. The average ratio of transrional/transversional nucleotide substitution of pairwise comparison among six species (including Anemia salina) was 1.200 $\pm$ 0.310 when whole region alas examined. However, the ratio showed some differences when the conservative regions and variable regions frere separatelv examined. The molecular phylogenies of the five species were constructed by using two different tree making methods. In general the results support the previously reported molecular phylogeny of the decapod crustaceans. However, our results indicate thats in the analysis of the sequence dat3, the UPGMA clustering method of the distance matrix method should be carefully employed after considering the rate of nucneotide substitution in the different regions of the molecule.

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An Introduction to Microsatellite Development and Analysis (Microsatellite 개발 및 분석법에 대한 소개)

  • Yun Young-Eun;Yu Jeong-Nam;Lee Byoung-Yoon;Kwak Myounghai
    • Korean Journal of Plant Taxonomy
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    • v.41 no.4
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    • pp.299-314
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    • 2011
  • The choice of molecular markers is the first step when selecting experimental plans in the field of population genetics. The popular molecular markers in population genetic studies are mainly allozyme, RAPD, RFLP, AFLP, microsatellite, SNP and ISSR. Among these, microsatellites are frequently found in nuclear, chloroplast and mitochondrial genome, showing a high level of polymorphism and nuclear microsatellites are codominant. Thus, it is a favorable molecular marker for population structure analyses and genetic diversity studies. Microsatellites are composed of tandem repeated 1~6 base pair nucleotide motifs and can be easily amplified by PCR reactions using locus specific primers. Because microsatellites have low cross-species transferability, however, they are only applicable between phylogenetically close species. In wild plants, the lack of genomic information and the high development cost of the microsatellite obstruct the wider use of microsatellites in plant population genetics research. In this review, we introduce the basis for microsatellite markers, the development process, and analytical methods as well as evolutionary models and their applications. In addition, possible genotyping errors which lead to erroneous conclusions are discussed.

Isolation and identification of Aureobasidium spp. from flowers of the Jeolla-do province in Korea (호남 지역 꽃으로부터 야생효모 Aureobasidium속 분리 및 동정)

  • Kim, Jeong-Seon;Lee, Miran;Song, Mi Young;Kwon, Soon-Wo;Kim, Soo-Jin;Hong, Seung-Beom;Park, Byeong-Yong;Yun, Bong Sik
    • The Korean Journal of Mycology
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    • v.46 no.4
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    • pp.415-425
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    • 2018
  • To study the characteristics of yeasts, 433 strains of the genus Aureobasidium were isolated from the flowers collected from Jeolla-do in Korea, and the diversity of the strains was confirmed through molecular phylogenetic and morphological analyses. Based on phylogenetic analysis of LSU rDNA seguences, the Aureobasidium strains from the Jeolla-do province were classified into six groups. The dominant species of flower-derived yeasts were Groups A and D. Since Groups B, E, and F were found only in Jeollanam-do, we can infer that the Aureobasidium is distributed more widely in Jeollanam-do than in the Jeollabuk-do province. Through LSU and ITS rDNA sequence analyses, Group A was identified as A. pullulans, Group B as A. melanogenum, and Group F as a putative new species of Aureobasidium. Groups C, D, and E do not completely match with A. leucospermi, A. namibiae, or A. subglaciale by LSU or ITS rDNA analysis but are closely related to those species. Comparisons of colony morphology are likely to be more helpful in distinguishing Groups C and D. The results of this study can provide useful characteristics for future studies of the genus Aureobasidium.

Stable expression of brazzein protein, a new type of alternative sweetener in transgenic rice (형질전환 벼에서 brazzein 감미단백질의 안정적인 발현)

  • Lee, Ye Rim;Akter, Shahina;Lee, In Hye;Jung, Yeo Jin;Park, So Young;Cho, Yong-Gu;Kang, Kwon Kyoo;Jung, Yu Jin
    • Journal of Plant Biotechnology
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    • v.45 no.1
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    • pp.63-70
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    • 2018
  • Brazzein is the smallest sweet protein and was isolated from the fruit pulp of Pentadiplandra brazzeana Baillon, native to tropical Africa. From ancient times, the indigenous people used this fruit in their diet to add sweetness to their daily food. Brazzein is 500 to 2000 times sweeter than sucrose on a weight basis and 9500 times sweeter on a molar basis. This unique property has led to increasing interest in this protein. However, it is expensive and difficult to produce brazzein other than in its native growing conditions which limits its availability for use as a food additive. In this study, we report high production yields of, brazzein protein in transgenic rice plants. An ORF region encoding brazzein and driven by the $2{\times}CaMV\;35S$ promoter was introduced into rice genome (Oryza sativa Japonica) via Agrobacterium-mediated transformation. After transformation, 17 regenerated plant lines were obtained and these transgene-containing plants were confirmed by PCR analysis. In addition, the selected plant lines were analyzed by Taqman PCR and results showed that 9 T0 lines were found to have a single copy out of 17 transgenic plants. Moreover, high and genetically stable expression of brazzein was confirmed by western blot analysis. These results demonstrate that recombinant brazzein was efficiently expressed in transgenic rice plants, and that we have developed a new rice variety with a natural sweetener.

Plant growth-promoting activity and identification of endophytic fungi isolated from native plant in East coast (동해안 자생식물로부터 분리된 내생균류의 식물생장촉진활성 및 동정)

  • You, Young-Hyun;Jin, Yong Ju;Kang, Sang-Mo;Oh, Sejong;Lee, Myung-Chul;Kim, Jong-Guk
    • Korean Journal of Microbiology
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    • v.51 no.1
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    • pp.14-20
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    • 2015
  • Coastal plant species, Plantago camtschatica Cham. native to the coastal region of the East Sea were sampled and then morphologically different 20 endophytic fungal strains were purely isolated. Phylogenetic analysis of isolates was done by the Bayesian program based on sequenced internal transcribed spacer (ITS-rDNA) region. Culture filtrates of each of 20 isolates were treated to Waito-c rice (WR) seedlings for verifying plant growth-promoting activity, respectively. As the results, E/PC/10/1 strain showed the highest plant growth-promoting activity among them. The culture filtrate of the strain E/PC/10/1 was revealed as containing gibberellins ($GA_1$, $GA_3$, $GA_4$) by using HPLC, and gas GC/MS with selected ion monitoring (SIM). Finally, this strain was identified as novel Penicillium spinulosum species that producing new GAs with microscopic observation and further molecular analysis with beta-tubulin gene sequence.

Identification of Arbuscular Mycorrhizal Fungi Colonizing Panax ginseng Using 18S rDNA Sequence (18S rDNA를 이용한 인삼(Panax ginseng)의 내생균근 균의 동정)

  • Eo, Ju-Kyeong;Kim, Dong-Hun;Jeong, Hyeon-Suk;Eom, Ahn-Heum
    • Applied Biological Chemistry
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    • v.47 no.2
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    • pp.182-186
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    • 2004
  • Morphological observation of roots and molecular technique were used to investigate the symbiotic relationships between arbuscular mycorrhizal (AM) fungi and ginseng roots. Korean ginseng, Panax ginseng, was collected from 8 sites in Korea. Colonization pattern of AM fungi in ginseng roots was determined as an Arum type under light microscopes. Nested PCR using AM fungal specific primers was employed to amplify a partial region on 18s rDNA of AM fungi from the root extracted mixed DNA. The amplified DNA was cloned and analyzed by random fragment length polymorphism (RFLP) with restriction enzymes, AluI, HinfI and AsuC21. One from each RFLP pattern was selected for sequencing. A total 16 clones were sequenced and identified as 2 species of AM fungi; Paraglomus brasilianum and Glomus spurcum. Paramglomus brasilianum was found from most of the ginseng roots, in this syudy suggesting that this species of AM fungi could have specific relationship with the ginseng root. Possible roles of AM fungal species in ginseng roots are discussed.

Animal species identification by co-amplification of hypervariable region 1 (HV1) and cytochrome b in mitochondrial DNA

  • Lim, Si Keun;Park, Ki Won
    • Analytical Science and Technology
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    • v.18 no.3
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    • pp.257-262
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    • 2005
  • Mitochondrial DNA (mt DNA) sequence analysis has been a useful tool for species identification of animals and human individuals. Two hypervariable regions (HV1 and HV2) in control region of mitochondrial genome were analyzed for human individual identification. In case of animal species identification, several genes on mt DNA such as cytochrome b (cytb), RNAs, cytochrome oxidases (CO) were used. In this study, co-amplification of HV1 and cytb was carried out in order to check the contamination of animal DNA and to verify the human DNA. The primer sets used in PCR were H15997/L16236 for HV1 and H14724/L15149 for cytb. PCR products for HV1 and cytb were 239 bp and 425 bp, respectively. The appearance of two bands on agarose gel implied the DNA came from human, however the single band of cytb gene represented the non-human animal DNA.

Analysis of genetic divergence according to each mitochondrial DNA region of Haliotis discus hannai (북방전복 (Haliotis discus hannai) 의 mitochondrial DNA 영역별 유전적 변이성 분석)

  • Park, Choul-Ji;Nam, Won Sick;Lee, Jeong-Ho;Noh, Jae Koo;Kim, Hyun Chul;Park, Jong Won;Hwang, In Jun;Kim, Sung Yeon
    • The Korean Journal of Malacology
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    • v.29 no.4
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    • pp.335-341
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    • 2013
  • The seven mitochondrial DNA regions (ND2, ND5, ND4, ND4L, ND6, ND1 and 12SrRNA) of Haliotis discus hannai were examined to estimate the availability as a genetic marker for the study of population genetic. The region with the highest genetic variation was ND4 (Haplotype diversity = 1.0000, Nucleotide diversity = 0.0108). On the other hand, ND2 and ND1 regions have significantly appeared genetic divergence between clusters (divergence of 90% and 87%). Also, pairwise $F_{ST}$ between clusters within ND2 and ND1 regions showed high values; 0.4061 (P = 0.0000), 0.4805 (P = 0.0000) respectively. Therefore we can infer that it is the most efficient and accurate way to analyze the region of ND4 with the highest variation in addition to the regions of ND2 and ND1, which formed clusters with high bootstrap value, for study of population genetic structure in this species.