A novel radiation-resistant Filobasidium sp. yeast strain was isolated from seawater. Along with this strain, a total of 656 yeast isolates were purified from seawater samples collected from three locations in the West Sea of Korea and assessed for their radiation tolerance. Among these isolates, fi...
A novel radiation-resistant Filobasidium sp. yeast strain was isolated from seawater. Along with this strain, a total of 656 yeast isolates were purified from seawater samples collected from three locations in the West Sea of Korea and assessed for their radiation tolerance. Among these isolates, five were found to survive a 5 kGy radiation dose. The most radiation-resistant strain was classified as Filobasidium sp. based on 18S rDNA sequence analysis and hence was named Filobasidium RRY1 (Radiation-Resistant Yeast 1). RRY1 differed from F. elegans, which is closely related to RRY1, in terms of the optimal growth temperature and radiation resistance, and was resistant to high doses of ${\gamma}$-ionizing radiation ($D_{10}$: 6-7 kGy). When exposed to a high dose of 3 kGy irradiation, the RRY1 cells remained intact and undistorted, with negligible cell death. When these irradiated cells were allowed to recover, the cells fully repaired their genomic DNA within 3 h of growth recovery. This is the first report in which a radiation-resistant response has been investigated at the physiological, morphological, and molecular levels in a strain of Filobasidium sp.
A novel radiation-resistant Filobasidium sp. yeast strain was isolated from seawater. Along with this strain, a total of 656 yeast isolates were purified from seawater samples collected from three locations in the West Sea of Korea and assessed for their radiation tolerance. Among these isolates, five were found to survive a 5 kGy radiation dose. The most radiation-resistant strain was classified as Filobasidium sp. based on 18S rDNA sequence analysis and hence was named Filobasidium RRY1 (Radiation-Resistant Yeast 1). RRY1 differed from F. elegans, which is closely related to RRY1, in terms of the optimal growth temperature and radiation resistance, and was resistant to high doses of ${\gamma}$-ionizing radiation ($D_{10}$: 6-7 kGy). When exposed to a high dose of 3 kGy irradiation, the RRY1 cells remained intact and undistorted, with negligible cell death. When these irradiated cells were allowed to recover, the cells fully repaired their genomic DNA within 3 h of growth recovery. This is the first report in which a radiation-resistant response has been investigated at the physiological, morphological, and molecular levels in a strain of Filobasidium sp.
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제안 방법
The cellular fatty acid (CFA) composition of the RRY1 strain was determined using a standard protocol of the MIDI/Hewlett Packard Microbial Identification System [1], in which fatty acid methyl esters of freeze-dried yeast cells (unirradiated or irradiated at 3 kGy) were extracted after saponification and methylation, and then analyzed by gas chromatography. The Agilent 6890N GC system (Agilent Technologies Inc., DE, USA) with a methyl phenyl silicone fused silica capillary column (HP 19092B-102) and flame ionization detector (FID) (carrier gas: H2; initial temperature: 170℃; final temperature: 270℃; FID temperature:300℃; injection port temperature: 250℃) was used for the analysis.
The samples were then coated with gold–palladium and examined using a model JSM-30 scanning electron microscope (JEOL, Japan).
The cells were harvested by centrifugation and fixed with 2% (v/v) glutaraldehyde solution at RT for 4 h. The samples were washed with PBS and dehydrated using a graded ethanol series (30%, 50%, 70%, 80%, and 100%). The samples were then coated with gold–palladium and examined using a model JSM-30 scanning electron microscope (JEOL, Japan).
대상 데이터
Seawater samples were collected from three locations in the West Sea of Korea: Maehwa Island, Chilsan Island, and Doripo Beach, located in Shin’an, Yeonggwang, and Muju, respectively, in Jeollanam Province.
이론/모형
Fatty acid analysis. The cellular fatty acid (CFA) composition of the RRY1 strain was determined using a standard protocol of the MIDI/Hewlett Packard Microbial Identification System [1], in which fatty acid methyl esters of freeze-dried yeast cells (unirradiated or irradiated at 3 kGy) were extracted after saponification and methylation, and then analyzed by gas chromatography. The Agilent 6890N GC system (Agilent Technologies Inc.
Phylogenetic tree based on 18S rDNA sequence data of the RRY1 strain. The tree was drawn using the neighbor-joining method [20]. Bar: 2 substitutions per 1,000 bases.
성능/효과
Growth and survival of Filobasidium RRY1 was compared with that of S. cerevisiae SC7931 (Korean Culture for Type Collection, Korea) and F. elegans CBS7640 (CBS-KNAW Fungal Biodiversity Centre, The Netherlands). All strains were inoculated in YPD broth, and growth curves were studied until the stationary phase at 20℃ and 25℃.
elegans was demonstrated in the present study, but the radiation resistance phenotype in Filobasidium had not yet been reported earlier. Thus, to validate our findings, the radioresistant phenotype of RRY1 was compared with Saccharomyces cerevisiae, the widely known yeast model used for radiation resistance studies. RRY1 (D10: 5-7 kGy) showed better growth and higher radiation resistance than S.
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