Improved Methodology for Identification of Cryptomonads: Combining Light Microscopy and PCR Amplification
자연과학 > 생물
( Shuang Xia ) , ( Ying Yin Cheng ) , ( Huan Zhu ) , ( Guo Xiang Liu ) , ( Zheng Yu Hu )
한국미생물생명공학회(구 한국산업미생물학회)
Journal of Microbiology and Biotechnology 2013년, 제23권 제3호, 289~296페이지(총8페이지)
05212220.pdf [무료 PDF 뷰어 다운로드]
  • ※ 본 자료는 참고용 논문으로 수정 및 텍스트 복사가 되지 않습니다.
  • 구매가격
    135원 (구매자료 3% 적립)
    이메일 발송  스크랩 하기
    자료 다운로드  네이버 로그인
    Cryptomonads are unicellular, biflagellate algae. Generally, cryptomonad cells cannot be preserved well because of their fragile nature, and an improved methodology should be developed to identify cryptomonads from natural habitats. In this study, we tried using several cytological fixatives, including glutaraldehyde, formaldehyde, and their combinations to preserve field samples collected from various waters, and the currently used fixative, Lugol`s solution was tested for comparison. Results showed that among the fixatives tested, glutaraldehyde preserved the samples best, and the optimal concentration of glutaraldehyde was 2%. The cell morphology was well preserved by glutaraldehyde. Cells kept their original color, volume, and shape, and important taxonomic features such as furrow/gullet complex, ejectosomes, as well as flagella could be observed clearly, whereas these organelles frequently disappeared in Lugol`s solution preserved samples. The osmotic adjustments and buffers tested could not preserve cell density significantly higher. Statistical calculation showed the cell density in the samples preserved by 2% glutaraldehyde remained stable after 43 days of the fixation procedure. In addition, DNA was extracted from glutaraldehyde preserved samples by grinding with liquid nitrogen and the 18S rDNA sequence was amplified by PCR. The sequence was virtually identical to the reference sequence, and phylogenetic analyses showed very close relationship between it and sequences from the same organism. To sum up, the present study demonstrated that 2% unbuffered glutaraldehyde, without osmotic adjustments, can preserve cryptomonads cells for identification, in terms of both light microscopy and phylogenetic analyses based on DNA sequences.
    오늘 본 자료
    • 오늘 본 자료가 없습니다.
    본 학술논문은 한국학술정보㈜ 각 학회간에 저작권 계약이 체결된 것으로 HAPPY학술이 제공하고 있습니다.
    본 저작물을 불법적으로 이용시는 법적인 제재가 가해질 수 있습니다.
    사업자등록번호 220-87-87785 대표.신현웅 주소.서울시 서초구 방배로10길 18, 402호 대표전화.070-8809-9397
    개인정보책임자.박정아 통신판매업신고번호 제2017-서울서초-1765호 이메일
    copyright (c) 2009 happynlife. steel All reserved.