Journal of Plant Biotechnology

  • 제31권3호
  • /
  • Pages.225-230
  • /
  • 2004
  • /
  • 1229-2818(pISSN)
  • /
  • 2384-1397(eISSN)
한국식물생명공학회 (The Korean Society of Plant Biotechnology)
권호 표지
DOI QR코드

DOI QR Code

Comet 분석을 통한 방사선처리 고추세포의 핵 DNA 손상평가

Assessment of Nucleus-DNA Damage in Red Pepper Cells Treated with γ-Radiation through Comet Assay

  • 발행 : 2004.09.01
PDF 다운로드
⟨ 이전 논문 다음 논문 ⟩

초록

방사선에 노출된 고추유묘의 잎으로부터 세포핵을 분리하여 단세포전기영동방법인 comet 분석을 통하여 핵 DNA의 손상정도를 조사하였다. Comet 분석에서 꼬리부분의 길이 (T)와 머리부분의 길이 (H)를 측정하여 T/H 비율을 조사하였다. 무처리세포는 T/H 비율이 1.28이었으나 50 및 100 Gy의 방사선을 처리한 세포는 각각 3.54 및 3.39로 방사선처리에 의해 상당량의 핵 DNA가 손상을 입은 것으로 나타났다. Comet의 head-DNA량은 무처리가 76.8%였으나 50 및 100 Gy를 처리한 세포는 각각 55.9% 및 59.5%를 보였다. 고선량의 방사선을 처리하기 전에 미리 20 Gy 이하의 저선량 방사선을 종자에 전처리하였을 경우 종자의 발아 및 생장에 대한 영향은 없었지만, 후속 고선량에 대한 핵 DNA의 손상은 경감되는 경향을 보였다.

We employed single cell gel electrophoresis method (comet assay) to analyze the degree of nucleus-DNA damage in the leaves of red pepper (Capsicum annuum L.) seedlings exposed to $^{60}$ CO v-radiation stress. Nucleus-DNA damage was measured as the ratio of tail length (T) to head length (H) in individual comet image isolated from pepper leaf cell. The T/H ratio of control-cells and treated-cells at 50 or 100 Gy were 1.28 and 3.54 or 3.39, respectively, suggesting that nuclei of pepper cells were severely damaged in the integrity of DNA strand by the treatment of enhanced v-radiation. The percentage of head-DNA in control-cells was 76.8%, whereas those of 50 and 100 Gy treated-cells were 55.9% and 59.9%, respectively. Pretreatment of low dose (4 to 20 Gy) radiation to seeds decreased DNA-damage in the leaves of seedlings treated with high dose radiation at 50 or 100 Gy. In this experiment, we developed a sensitive, reliable and rapid method for evaluating genotoxic effect in the nuclei of plant cells by employing comet assay.

키워드

참고문헌

  1. Atkisnson GF (1989) Report upon some preliminary experiment with the Rontgen rays on plant. Science 7: 7-9
  2. Fairbairn DW, Olivie PL, O' Neill K (1995) The comet assay: a comprehensive review. Mutation Res 339: 37-44 https://doi.org/10.1016/0165-1110(94)00013-3
  3. Gichner T, Plewa MJ (1998) Induction of somatic DNA damage as measured by single cell gel electrophoresis and point mutation in leaves of tobacco plants. Mutation Res 401: 143-152 https://doi.org/10.1016/S0027-5107(98)00003-7
  4. Gichner T, Ptacek O, Stavreva DA, Wagner ED, Plewa MJ (2000) A comparison of DNA repair using the comet assay in tobacco seedlings after exposure to alkylating agents or ionizing radiation. Mutagenesis 15: 503-506 https://doi.org/10.1093/mutage/15.6.503
  5. Kim JS, Lee YK, Song HS, Park HS, Kim JK (1999) Effects of low dose ionizing radiation on the growth and yield of soybean cultivars. Kor J Environ Agriculture 18: 66-69
  6. Koppen G, Angelis KJ (1998) Repair of X-ray induced DNA damage measured by the comet assay in roots of Vicia faba. Environ Mol Mutagen 32: 281-285 https://doi.org/10.1002/(SICI)1098-2280(1998)32:3<281::AID-EM11>3.0.CO;2-R
  7. Koppen G, Verschaeve L (1996) The alkaline comet test on plant cells, a new genotoxicity test for DNA strand breaks in Vicia faba root cells. Mutation Res 360: 193-200 https://doi.org/10.1016/S0165-1161(96)90017-5
  8. Kwon ST, Jung EA, Kim JS (2001) Effect of γ-radiation on growth and antioxidant enzyme activities in red pepper. Kor J Life Science 11: 612-617
  9. Menke M, Meister A, Schubert I (2000) N-methyl-N-nitrosourea- induced DNA damage detected by the comet assay in Vicia faba nuclei during all interphase stages is not restricted to chromatid aberration hot spots. Mutagenesis 15: 503-506 https://doi.org/10.1093/mutage/15.6.503
  10. Miller MW, Miller WM (1987) Radiation hormeosis in plants. Health Physics 52: 607-616 https://doi.org/10.1097/00004032-198705000-00012
  11. Navarrette MH, Carrera P, Mitguel M, Torre C (1997) A fast comet assay variant for solid tissue cells. The assessment of DNA damage in higher plants. Mutation Res 389: 271-279 https://doi.org/10.1016/S1383-5718(96)00157-X
  12. Noroozi, M, Angerson WJ, Lean MEJ (1998) Effects of flavonoids and vitamin C on oxidative DNA damage to human lymphocytes. Am J Clinical Nutrients 64: 1210-1218
  13. Rojas E, Lopez MC, Valverde M (1999) Single cell gel electrophoresis assay; methodology and application. J Chromatography B 772: 225-254
  14. Sheppard SC, Evenden WG (1986) Factors controlling the hormeosis response of field crops to very low does of gamma irradiation of seed. Can J Plant Sci 66: 431-435 https://doi.org/10.4141/cjps86-061
  15. Stan S, Croitoru A (1970) Effect of low, moderate and high levels of gamma radiations on soybean plants. I. Analysis of growth and yield. Stim Newsl 1: 23-25
  16. Stepleton AE, Walbot V (1994) Flavoniods can protect maize DNA from the induction of ultraviolet radiation damage. Plant Physiol 105: 881-889 https://doi.org/10.1104/pp.105.3.881