• ALGAE
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• 제17권3호
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• pp.153-159
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• 2002

Partial 16S rRNA gene sequences of seven cyanophycean strains from the National Instiute of Environmental Research of Korea - Microcystis aeruginosa, M. aeruginosa f. aeruginosa, M. ichthyoblade, M. viridis, Anabaena flos-aquae, and Oscillatoria sancta - were analyzed and the phylogenetic relationship of Microcystis among Cyanophyceae were evaluated. Based on sequence analysis results, Microcystis is monophyletic, the clade of which supported 100% bootstrap tress, and distinguished clearly from the other taxa. Therefore, the partial 16S rRNA gene sequences can be a useful and efficient tool for distinguishing Microcystis from other cyanophycean without axenic culture or cloning.

• 한국해양바이오학회지
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• 제2권2호
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• pp.77-80
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• 2007

Macroalgal tissues can be used as indicating materials for environmental assessment using several algal biotechnology techniques. As bioassay test organisms, macroalgal tissues are required as an axenic state for suitable biological indicators. Callus formation and blade regeneration under suitable culture conditions are also useful for the tests. Quantitative method using tetrazolium chloride or $alamarBlue^{TM}$ is devised on a rapid assessment of the seaweed viability. The use of RT-PCR especially differential display technique should provide the means for the detection and isolation of the responding genes induced by the environmental stress. Seaweed thriving in more environmental changes might contain more diverse biologically active substances.

• Mycobiology
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• 제32권4호
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• pp.197-198
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• 2004

A new technique for single spore isolation was developed for predacious fungi forming constricting rings directly on the spores using Dactylaria brochopaga and Arthrobotrys dactyloides. Constricting rings were induced directly on the spores by transferring the spores in 25 ppm solution of DL-Valine in sterile distilled water. Freshly hatched and thoroughly washed second stage juveniles of Meloidogyne incognita were transferred into cavity blocks containing induced rings for trapping and killing of nematodes. The killed nematodes were surface sterilized with streptomycin and inoculated into petri dishes containing maize meal agar media with 100 ppm streptomycin. The petri dishes were incubated at $29{\pm}1^{\circ}C$ for few days which yielded axenic culture of these fungi.

• Journal of Animal Science and Technology
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• 제65권2호
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• pp.387-400
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• 2023

Ruminal protozoa, especially entodiniomorphs, engulf other members of the rumen microbiome in large numbers; and they release oligopeptides and amino acids, which can be fermented to ammonia and volatile fatty acids (VFAs) by amino acid-fermenting bacteria (AAFB). Studies using defaunated (protozoa-free) sheep have demonstrated that ruminal protozoa considerably increase intraruminal nitrogen recycling but decrease nitrogen utilization efficiency in ruminants. However, direct interactions between ruminal protozoa and AAFB have not been demonstrated because of their inability to establish axenic cultures of any ruminal protozoan. Thus, this study was performed to evaluate the interaction between Entodinium caudatum, which is the most predominant rumen ciliate species, and an AAFB consortium in terms of feed degradation and ammonia production along with the microbial population shift of select bacterial species (Prevotella ruminicola, Clostridium aminophilum, and Peptostreptococcus anaerobius). From an Ent. caudatum culture that had been maintained by daily feeding and transfers every 3 or 4 days, the bacteria and methanogens loosely associated with Ent. caudatum cells were removed by filtration and washing. An AAFB consortium was established by repeated transfers and enrichment with casamino acids as the sole substrate. The cultures of Ent. caudatum alone (Ec) and AAFB alone (AAFB) and the co-culture of Ent. caudatum and AAFB (Ec + AAFB) were set up in three replicates and incubated at 39℃ for 72 h. The digestibility of dry matter (DM) and fiber (NDF), VFA profiles, ammonia concentrations, pH, and microscopic counts of Ent. caudatum were compared among the three cultures. The co-culture of AAFB and Ent. caudatum enhanced DM degradation, VFA production, and Ent. caudatum cell counts; conversely, it decreased acetate: propionate ratio although the total bacterial abundance was similar between Ec and the Ec + AAFB co-culture after 24 h incubation. The ammonia production and relative abundance of C. aminophilum and P. anaerobius did not differ between AAFB alone and the Ec + AAFB co-culture. Our results indicate that Ent. caudatum and AAFB could have a mutualistic interaction that benefited each other, but their interactions were complex and might not increase ammoniagenesis. Further research should examine how such interactions affect the population dynamics of AAFB.

• ALGAE
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• 제24권3호
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• pp.163-168
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• 2009

Grateloupia filicina (J.V. Lamouroux) C. Agardh (Halymeniaceae, Cryptonemiales, Rhodophyta) is an edible seaweed as well as an important source of carrageenan. In the present study, attempt has been made to develop a suitable protocol for effective regeneration of the seaweed and the rapid multiplication of the desired varieties. The young upright thallus of G. filicina was grown in axenic culture using both solid and liquid media. The various media tested were f/2, Provasoli’s Enriched Seawater (PES) and Enriched Seawater (ESW). The effect of glycerol (as a carbon source) and various plant growth regulators i.e. auxin (NAA) and cytokinins (Kinetin and BA) were tested. Although, regeneration of young thalli was observed from the cut ends in all the media, better growth was found in f/2, PES, f/2 (0.5% Glycerol), f/2 (NAA ${10^{-5}}_M)\;and\;f/2\;(BA\;{10^{-6}}_M$). On the other hand callusing was observed only in solid media supplemented with low concentration of Glycerol (0.5%) in f/2, NAA ${10^{-5}}_M\;in\;f/2,\;PES\;and\;BA\;{10^{-5}}_M$ in f/2. Young thalli were developed from the callus sub culture after 40 days of inoculation.

• 한국토양비료학회지
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• 제20권3호
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• pp.269-274
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• 1987

간척지(干拓地) 토양(土壤)의 잡초(雜草) 및 수도근(水稻根) 조직(組織)에서 분리동정(分離同定)한 수종(數種)의 협생질소고정균(協生窒素固定菌)을 질소(窒素)를 함유(含有)하지 않은 순수배양(純粹培養) 내(內)에서 재배형(栽培型) 및 품종(品種)이 상이(相異)한 수도종자권(水稻種子圈)에 접종(接種)하여 수도(水稻)의 건물중(乾物重), 초장(草長), 근장(根長), 근수등(根數等)의 생육특징(生育特徵)과 협생질소고정력(協生窒素固定力)을 조사(調査)한 결과(結果) 수도품종(水稻品種) 및 협생질소고정균(協生窒素固定菌)의 조합간(組合間) 교호작용(交互作用)은 통계적(統計的)으로 유의(有意)한 상관관계(相關關係)를 보이지 않했다. 그러나 공시수도품종중(供試水稻品種中)에서 Annapuruna는 7종(種)의 협생질소고정균(協生窒素固定菌)을 접종(接種)하므로써 생육량(生育量) 증가(增加)를 보였다. 그리고 7종(種)의 협생질소고정균(協生窒素固定菌)을 교호접종(交互接種)했을때 협생질소고정력(協生窒素固定力)은 Annapuruna와 신광(新光)벼 조합(組合)에서 가장 높았으며 IR-8이 가장 낮았다. 또한 협생질소고정균(協生窒素固定菌)과 수도품종간(水稻品種間)에는 신광(新光)벼와 Pseudomonas sp. H8 조합(組合)에서 질소고정력(窒素固定力)이 가장 높았다.

• 한국양식학회지
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• 제2권1호
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• pp.1-7
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• 1989

양식 해조류인 큰방사무늬김에 대한 조직배양을 종자 세포 보존 및 장차 세포수준의 내저 염성 품종개량을 목적으로 시도하였다. 김 엽체의 영양 성장부위를 무균적으로 PES-한천배지 상에 배양함으로서 callus의 형성을 유도하였으며 이 적갈색의 연약한 callus를 $16^{\circ}C$에서 12시간 주기로 2000 lux의 형광등 하에서 1개월마다 이식하면서 배양하였다. 총 탄수화물 및 단백질 함량을 측정하였으며, 최적 배양온도는 $14^{\circ}C\~18^{\circ}C$였다. PES-한천배지 상에서 $2.0\%$의 NaCl 농도가 callus의 성장에 가장 우수하였다.

• Journal of Microbiology and Biotechnology
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• 제10권3호
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• pp.429-433
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• 2000

Giardia lamblia, a human pathogen causing outbreaks of diarrhea, recently became a focus of great concerns in the fields of both medical and environmental microbioloty. To develop the experimental tools to study giardiasis, encystation, one of the major processes in its life cycle, was reconstituted by inducing an axenic culture of a flagellated form of G. lamblia into a cyst from under high concentration of bile and alkaline pH condition. The successful induction was confirmed by Northern analysis of resulting increased expression of the CWPl gene encoding the cyst wall protein 1. An examination of the encystation process with SEM (scanning electron microscopy) and TEM (transmission electron microscopy) revealed that the trophozoite, a flagellate with a bilateral symmetry, was transformed to a cyst form with an oval-shape and defined filamentous wall. The encystation was found to cause a disappearance of the flagella and an invagination of the adhesive disc. An extensive formation of rER (rough endoplasmic reticulum) was observed after 24h of induction, indication an active synthesis and export of proteins during this process. The vital staining of the invitro-induced systs showed that most cysts maintained their viability.

• Parasites, Hosts and Diseases
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• 제55권1호
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• pp.81-84
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• 2017

Trypanosoma cruzi is the etiological agent of Chagas disease. Epimastigote forms of T. cruzi can be readily cultured in axenic conditions. Ethanol and dimethyl sulfoxide (DMSO) are commonly used solvents employed as vehicles for hydrophobic compounds. In order to produce a reference plot of solvent dependent growth inhibition for T. cruzi research, the growth of epimastigotes was analyzed in the presence of different concentrations of ethanol (0.1-4.0%) and DMSO (0.5-7.5%). The ability of the parasites to resume growth after removal of these solvents was also examined. As expected, both ethanol and DMSO produced a dose-dependent inhibition of cellular growth. Parasites could recover normal growth after 9 days in up to 2% ethanol or 5% DMSO. Since DMSO was better tolerated than ethanol, it is thus recommended to prefer DMSO over ethanol in the case of a similar solubility of a given compound.

• The Plant Pathology Journal
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• 제19권3호
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• pp.152-158
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• 2003

Stromata of the white root rot fungus, Rosellinia necatrix, were produced on diseased roots although they were reported to develop rarely in nature. Forty-two (42) out of 47 samples produced synnemata while 23 developed stromata. Forty-seven (47) isolates obtained from diseased root samples were divided into 24 mycelium compatibility groups (MCGs). Sixteen (16) out of 24 MCGs produced stromata. Single ascospore isolates from 10 stroma samples produced dsRNA-containing isolates from diseased tissue beneath stromata. The frequency of synnema production on axenic culture varied among isolates with different origin. The dsRNA was not transmitted vertically to the ascospore offspring despite the infection of various dsRNA in the parental isolates. The dsRNA was absent in 35 ascospore isolates in two stroma samples that originated from the isolates, in which dsRNA was not eliminated by hyphal tip isolation. Consequently, sexual reproduction in the white root rot fungus was suggested to produce propagules as a new infection source and to have the function to eliminate infectious factors such as mycoviruses.