• Journal of Plant Biology
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• 제35권2호
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• pp.99-106
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• 1992

인삼(Panax ginseng C.A. Meyer) 종자단백질인 vicilin을 ammonium sulfate 침전, gel permeation 및 이온 교환 크로마토그래피로 정제하였다. Vicilin은 분자량 55,000(큰 소단위) 및 44,000(작은 소단위)인 두 종의 소단위를 포함하는 당단백질이다. Vicilin에 대한 항체를 토끼에서 형성시켜 DEAE-Affi-Gel Blue affinity 크로마토그래피로 정제하였다. 이 항체와 금 입자가 결합된 2차 항체를 종자의 배유세포에 반응시켰다. 금 입자는 배유세포내의 단백질체, 전자밀도가 높은 과립 및 골지체의 elaborating 과립에 표지되었다. 이러한 결과는 조면소포체에서 합성되어 골지체로 수송된 vicilin이 골지의 소포내에서 공정과정을 거쳐 전자밀도가 높은 과립이 된 다음 단백질로 수송됨을 나타낸다.

• ALGAE
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• 제31권3호
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• pp.219-230
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• 2016

The Euglena deses group are common freshwater species composed of E. adhaerens, E. carterae, E. deses, E. mutabilis, and E. satelles. These species are characterized by elongated cylindrical worm-like cell bodies and numerous discoid chloroplasts with a naked pyrenoid. To understand the cryptic diversity, species delimitation and phylogenetic relationships among members of the group, we analyzed morphological data (light and scanning electron microscopy) and molecular data (nuclear small subunit [SSU] and large subunit [LSU] rDNAs and plastid SSU and LSU rDNAs). Bayesian and maximum likelihood analyses based on the combined four-gene dataset resulted in a tree consisting of two major clades within the group. The first clade was composed of two subclades: the E. mutabilis subclade, and the E. satelles, E. carterae, and E. adhaerens subclade. The E. mutabilis subclade was characterized by a lateral canal opening at the anterior end and a single pellicular stria, whereas the E. satelles, E. carterae, and E. adhaerens subclade was characterized by an apical canal opening at the anterior end of the cell and double pellicular striae. The second clade consisted of 20 strains of E. deses, characterizing by a subapical canal opening at the anterior end and double pellicular striae, but they showed cell size variation and high genetic diversity. Species boundaries were tested using a Bayesian multi-locus species delimitation method, resulting in the recognition of five cryptic species within E. deses clade.

• Journal of Microbiology and Biotechnology
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• 제5권4호
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• pp.194-199
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• 1995

A portion (7.4 kb) of ribosomal DNA tandem repeat unit from a genome of the mushroom T. matsutake has been cloned. A 1.75 kb EcoRI fragment was cloned first using S. cerevisiae 255 rRNA gene as a probe, and this was then used for further cloning. A chromosomal walking experiment was carried out and the upstream region of the 1.75 kb fragment was cloned using SmaI/BamHI enzyme, the size was estimated to be 5.2 kb in length. Part of the downstream region of the 1.75 kb fragment was also cloned using XbaI/BamHI enzymes. Restriction enzyme maps of three cloned DNA fragments were constructed. Northern hybridization, using total RNA of T. matsutake, and the restriction fragments of three cloned DNAs as probes, revealed that all four ribosomal RNA genes (large subunit[LSU], small subunit [SSU], 5.85 and 5S rRNA genes) are present in the cloned region. The gene organization of the rDNA are regarded as an intergenic spacer [IGS]2 (partial) - SSU rRNA - internal transcribed spacer [ITS]1 - 5.8S rRNA - ITS2 - LSU rRNA - IGS1 -5S rRNA - IG52 (partial).

• ALGAE
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• 제35권2호
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• pp.157-165
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• 2020

Culture isolates of the genus Hypoglossum (Delesseriaceae, Rhodophyta) were obtained and their development and morphological structure over many years were followed in the laboratory. Molecular data (rbcL, large subunit ribosomal DNA, and cytochrome c oxidase subunit I) were obtained from these strains and evidence presented to recognize the new species: Hypoglossum sabahense from Sabah, Malaysia. Because various aspects of morphology in culture specimens differ significantly from types based on field specimens we have to rely mainly on the molecular criteria in ascribing a new taxonomic name here. This also is complicated by the major lack of molecular phylogenetic evidence for Hypoglossum and other Delesseriaceae. The 'Germling Emergence Method' and 'serendipity' are proving valuable in discovering significant new taxa from laboratory cultures which otherwise might never be known.

• Journal of Microbiology
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• 제43권3호
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• pp.266-271
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• 2005

In eukaryotic cells, various proteins are anchored to the plasma membrane through glycosylphosphatidylinositol (GPI). To study the biosynthetic pathways and modifications of GPI, various mutant cells have been isolated from the cells of Chinese hamster ovaries (CHO) supplemented with several exogenous genes involved in GPI biosynthesis using aerolysin, a toxin secreted from gram-negative bacterium Aeromonas hydrophila. Alpha toxin from Gram-positive bacterium Clostridium septicum is homologous to large lobes (LL) of aerolysin, binds GPI-anchored proteins and possesses a cell-destroying mechanism similar to aerolysin. Here, to determine whether alpha toxins can be used as an isolation tool of GPI-mutants, like aerolysin, CHO cells stably transfected with several exogenous genes involved in GPI biosynthesis were chemically mutagenized and cultured in a medium containing alpha toxins. We isolated six mutants highly resistant to alpha toxins and deficient in GPI biosynthesis. By genetic complementation, we determined that one mutant cell was defective of the second subunit of dolichol phosphate mannose synthase (DPM2) and other five cells were of a putative catalytic subunit of inositol acyltransferase (PIG-W). Therefore, C. septicum alpha toxins are a useful screening probe for the isolation of various GPI-mutant cells.

• Mycobiology
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• 제43권4호
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• pp.392-401
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• 2015

Using dilution plating method, 47 fungal isolates were obtained from a soil sample collected from Dokdo in the East Sea of Korea in 2013. In this study, two fungal isolates, EML-MFS30-1 and EML-DDSF4, were confirmed as undescribed species, Metarhizium guizhouense and Mortierella oligospora in Korea based on current classification system using multi loci including rDNA internal transcribed spacer, large subunit, small subunit, and ${\beta}$-tubulin (BTUB) genes. Herein, detailed morphological descriptions on characters of the undescribed fungal species as well as their molecular phylogenetic status are provided with comparisons to related species.

• 한국균학회지
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• 제48권3호
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• pp.187-195
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• 2020

Two unreported fungal isolates, KNU-GW1901 and KNU-AP100C, were collected from soil sample in Gyeongsangbuk-do, Korea. Their cultural and morphological characteristics were examined after 4 weeks of incubation at 25℃ on potato dextrose agar (PDA), malt extract agar (MEA), and oatmeal agar (OA). The conidial shape of KNU-GW1901 was aseptate, hyaline, globose to ellipsoidal, oblong, and reniform to pyriform and 2.61-4.97×1.93-3.61 ㎛ in size, whereas no conidial structures were observed in KNU-AP100C. The internal transcribed spacer (ITS) regions, large subunit (LSU), and small subunit (SSU) sequences were used to determine the taxonomic positions of the strains using the maximum likelihood phylogenetic tree. The isolate KNU-GW1901 was closely clustered with Plenodomus sinensis MFLUCC 17-0767, and KNU-AP100C was closely matched with P. collinsoniae CBS 120227. Based on the findings of morphological, cultural, and phylogenetic analysis, the isolates KNU-GW1901 and KNU-AP100C were identical to the previously described P. sinensis and P. collinsoniae isolates, respectively, which are first reported in Korea.

• 한국연초학회지
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• 제17권1호
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• pp.20-26
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• 1995

This study was carried out to obtain the basic data which include the change of the photosynthetic rate and protein content according to growth stage in the process of senescence of tobacco plant The photosynthetic rate was the maximum with 26.31$\mu$mol.CO2/m2.sec and stomatal resistance was the minimum with 0.2552cm/sec at 15th days after leaf emergence. However, after 50 days the photosynthesis was very little occurred. During leaf developments the number of chloroplast was increased and reached at the maximum at 25th days after emergence of leaf, thereafter, it was decreased gradually. The content of protein increased continuously and showed the highest value at 15th days after leaf emergence. The degradation rate of soluble protein was more rapid than that of insoluble protein at early stage of senescence. The range of decrement in the insoluble protein was low at late stage of senescence. The content of Rubisco, the key enzyme of photoamthesis, corresponded to about 50% of soluble protein and reached to the maximum at 150 days after leaf emergence. As the senescence progressed, the content of large subunit(UV) of Rubisco showed a tendency to decrease more rapidly than that of small subunit(SSU). The total amount of amino acids was the highest at 15th days after leaf emergence.

• ALGAE
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• 제28권4호
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• pp.307-330
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• 2013

The genus Cryptomonas is easily recognized by having two flagella, green brownish color, and a swaying behavior. They have relatively simple morphology, and limited diagnostic characters, which present a major difficulty in differentiating between species of the genus. To understand species delineation and phylogenetic relationships among Cryptomonas species, the nuclear-encoded internal transcribed spacer 2 (ITS2), partial large subunit (LSU) and small subunit ribosomal DNA (rDNA), and chloroplast-encoded psbA and LSU rDNA sequences were determined and used for phylogenetic analyses, using Bayesian and maximum likelihood methods. In addition, nuclear-encoded ITS2 sequences were predicted to secondary structures, and were used to determine nine species and four unidentified species from 47 strains. Sequences of helix I, II, and IIIb in ITS2 secondary structure were very useful for the identification of Cryptomonas species. However, the helix IV was the most variable region across species in alignment. The phylogenetic tree showed that fourteen species were monophyletic. However, some strains of C. obovata had chloroplasts with pyrenoid while others were without pyrenoid, which used as a key character in few species. Therefore, classification systems depending solely on morphological characters are inadequate, and require the use of molecular data.

• Mycobiology
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• 제49권5호
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• pp.439-453
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• 2021

During the rainy season in Thailand, specimens of Phallus chiangmaiensis sp. nov. and P. merulinus were collected from Chiang Mai and Samut Sakhon Provinces, respectively. Molecular phylogenetic analyses based on sequences of the nuclear ribosomal large subunit (LSU), nuclear ribosomal 5.8S gene including the internal transcribed spacer regions 1 and 2 (ITS), and the protein-coding gene atp6 (mitochondrial adenosine triphosphate [ATP] synthase subunit 6) support the placement of the new species within Phallus. Phallus chiangmaiensis has a well-developed white indusium and campanulated caps with reticulate surfaces. It differs morphologically from the related species, as supported by the phylogenetic data. Phallus merulinus is reported here as a species that was re-encountered in Thailand. The descriptions of the species are accompanied by illustrations of macro- and micro- morphological features, and a discussion of the related taxa is presented.