Journal of the Korea Academia-Industrial cooperation Society (한국산학기술학회논문지)

The Korea Academia-Industrial cooperation Society (한국산학기술학회)
권호 표지
DOI QR코드

DOI QR Code

Cloning and Expression of Phytochelatin Synthase 1 Gene from Rhizophora stylosa Exposed to Cadmium and Copper

카드뮴과 구리에 노출된 Rhizophora stylosa 의 phytochelatin synthase 1 유전자 클로닝 및 발현

  • Lee, Gunsup (South Sea Environment Research Department, Korea Institute of Ocean Science and Technology) ;
  • Hwang, Jinik (South Sea Environment Research Department, Korea Institute of Ocean Science and Technology) ;
  • Park, Mirye (South Sea Environment Research Department, Korea Institute of Ocean Science and Technology) ;
  • Chung, Youngjae (Department of Life Science and Biotechnology, Shin Gyeong University) ;
  • Lee, Taek-Kyun (South Sea Environment Research Department, Korea Institute of Ocean Science and Technology)
  • 이건섭 (한국해양과학기술원 남해특성연구부) ;
  • 황진익 (한국해양과학기술원 남해특성연구부) ;
  • 박미례 (한국해양과학기술원 남해특성연구부) ;
  • 정영재 (신경대학교 생명공학과) ;
  • 이택견 (한국해양과학기술원 남해특성연구부)
  • Received : 2013.03.29
  • Accepted : 2013.06.07
  • Published : 2013.06.30
Download PDF
⟨ Previous Next ⟩

Abstract

The mangrove ecosystems have the capacity to act as a sink of heavy metals entering aquatic ecosystems. Despite their potential exposure to metal contaminated sediments, mangroves appear to be highly tolerant to heavy metals. In this study, we cloned metal tolerance gene from mangrove plant. Using CTAB method, RNA were isolated from leaves and root tissue of Rhizophora stylosa habitated at Weno island in Micronesia Chuuk lagoon using CTAB method and phytochelatin synthase 1 (PCS1) gene was cloned using gene specific primers. Expression of PCS1 gene was increased 1.91 fold and 2.72 fold in mangrove propagules exposed to 100 ppb Cd and 10 ppb Cu, respectively. These results indicate that expression of PCS1 gene are promising tools for health assessment of mangrove ecosystem.

망그로브 생태계는 수생태계로 유입되는 중금속을 받아들이는 기능을 가지고 있다. 중금속에 오염된 퇴적물에 노출됨에도 불구하고 망그로브는 중금속에 내성을 가지고 있다. 이 연구에서 우리는 망그로브로부터 중금속 저항성 관련 유전자를 클로닝하고, 중금속 노출에 유전자 발현 변화를 분석하였다. 미크로네시아 축라군의 웨노섬에서 채취한 Rhizophora stylosa의 잎과 뿌리조직으로부터 CTAB 방법을 이용하여 RNA를 분리하였고, gene specific primers를 이용하여 phytochelatin synthase 1(PCS1) 유전자를 클로닝하였다. R. stylosa 태생종자를 100 ppb의 Cd과 10 ppb의 Cu에 노출하였을 때 각각 1.91배 및 2.72배 발현이 증가하였다. 이러한 결과는 PCS1 유전자의 발현분석이 망그로브 생태계의 건강성을 평가하기 위한 좋은 도구가 될 수 있음을 나타낸다.

Keywords

References

  1. P. Saenger, D. McConchie, M. W. Clark, "Mangrove forests as a buffer zone between anthropologically polluted areas and the sea". In: Saenger, P. (Ed.), Proceedings of the 1990 CZM Workshop, Yeppoon, Qld, pp 280-297, 1990.
  2. E. C. Peters, N. J. Gassman, J. C. Firman, R. H. Richmond, E. A. Power, "Ecotoxicology of tropical marine ecosystems". Environ. Toxicol. Chem. vol. 16, no. 1, pp. 12-20, 1997. DOI: http://dx.doi.org/10.1002/etc.5620160103
  3. A. I. Roberson, M. J. Phillips, "Mangroves as filter of shrimp pond effluent: prediction and biogeochemical research needs". Hydrobiologia. vol. 295, no. 1-3, pp. 311-321, 1995. DOI: http://dx.doi.org/10.1007/BF00029138
  4. N. F. Y. Tam, Y. S. Wong, "Retention and distribution of heavy metals in mangrove soils receiving wastewater". Environ. Pollut. vol. 94, no. 3, pp. 283-291, 1996. DOI: http://dx.doi.org/10.1016/S0269-7491(96)00115-7
  5. J. Vangronsveld, H. Clijsters, "Toxic effects of metals". In: Farago, M.E. (Ed.), Plants and The Chemical Elements-Biochemistry, Uptake, Tolerance and Toxicity. VCH, Weinheim, pp. 149-177, 1994.
  6. S. Clemens, "Molecular mechanisms of plant metal tolerance and homeostasis". Planta vol. 212, no. 4, pp. 475-486, 2001. DOI: http://dx.doi.org10.1007/s004250000458
  7. J. L. Hall, "Cellular mechanisms for heavy metal detoxification and tolerance". J. Exp. Bot. vol. 53, no. 366, pp. 1-11, 2002. DOI: http://dx.doi.org/10.1093/jexbot/53.366.1
  8. W. E. Rauser, "Phytochelatins and related peptides: structure, biosynthesis, and function". Plant Physiol. vol. 109, no. 4, pp. 1141-1149, 1995. https://doi.org/10.1104/pp.109.4.1141
  9. C. Cobbett, P. Goldsbrough, "Phytochelatins and metallothioneins: roles in heavy metal detoxification and homeostasis". Annu. Rev. Plant Biol. vol. 53, pp. 159-182, 2002. DOI: http://dx.doi.org/10.1146/annurev.arplant.53.100301.135154
  10. G. R. MacFarlane, "Leaf biochemical parameters in Avicennia marina (Forsk.) Vierh as potential biomarkers of heavy metal stress in estuarine ecosystems". Mar. Pollut. Bull. vol. 44, no. 3, pp. 244-256, 2002. DOI: http://dx.doi.org/10.1016/S0025-326X(01)00255-7
  11. X. Fu, S. Deng, G. Su, Q. Zeng, S. Shi, "Isolating high-quality RNA from mangrove without liquid nitrogen". Plant Mol. Biol. Reporter, vol. 22, no. 2, pp. 197a-197e, 2004. DOI: http://dx.doi.org/10.1007/BF02772728
  12. W. D. Loomis, "Overcoming problems of phenolic and quinines in the isolation of plant enzymes and organelles". Meth. Enzymol. vol. 31, pp. 528-545, 1974. https://doi.org/10.1016/0076-6879(74)31057-9
  13. D. Gonzalez-Mendoza, A. Q. Moreno, O. Zapata-Perez, "Coordinated responses of phytochelatin synthase and metallothionein genes in black mangrove, Avicennia germinans, exposed to cadmium and copper". Aquatic Toxicol. vol. 83, no. 4, pp. 306-314, 2007. DOI: http://dx.doi.org/10.1016/j.aquatox.2007.05.005
  14. C. Harty, "Mangroves in New South Wales and Victoria". Vista Publications, Melbourne, pp. 47, 1997.
  15. R. J. West, C. A. Thorogood, R. J. Williams, "Environmental stress causing mangrove 'dieback' in NSW". Aust. Fisheries, vol. 42, no. 8, 16-20, 1983.
  16. O. J. Eong. "The ecology of mangrove conservation and management". Hydrobiologia, vol. 295, no, 1-3, pp. 343-351, 1995. DOI: http://dx.doi.org/10.1007/BF00029141
  17. D. E. Salt, R. D. Smith, I. Raskin, "Phytoremediation". Annu. Rev. Plant Physiol. Plant Mol. Biol. vol. 49, no. 1, pp. 643-668, 1998. DOI: http://dx.doi.org/10.1146/annurev.arplant.49.1.643
  18. D. Gleba, N. Y. Borisjuk, L. G. Borisjuk, R. Kneer, A. Poulev, M. Skarzhinskaya, S. Dushenkov, S. Logendra, Y. Y. Gleba, I. Raskin, "Use of plant roots for phytoremediation and molecular farming". Proc. Natl. Acad. Sci. USA vol. 96, no. 11, pp. 5973-5977, 1999. DOI: http://dx.doi.org/10.1073/pnas.96.11.5973
  19. F. Navari-Izzo, M. F. Quartacci, "Phytoremadiation of metals. Tolerance mechanisms against oxidative stress". Minerva Biotec. vol. 13, pp. 73-83, 2001