Selection of Cadmium Resistant Cell Line from Calli of Nicotiana tabacum cv. BY4

담배(Nicotiana tabacum L. cv. BY4)캘러스로부터 카드뮴 저항성 세포주의 선발

  • Published : 1997.11.01

Abstract

This study was carried out to select cadmium resistant cell lines from leaf-derived calli of diploid and haploid of Nicotiana tabacum cv. BY4, for understanding adaptation mechanism of plants in cadmium contaminated environment. suspended cell clumps were plated onto selection medium containing 0 to 2,000 $\mu$M cadmium. Cadmium resistant colonies were formed on the selection medium after 3 or 4 weeks of culture. The minimum inhibition concentration (MIC) of cadmium on colony formation were 300 $\mu$M in diploid and 200 $\mu$M in haploid plants, respectively. In order to test the resistance to cadmium, selected cell line on MIC were transferred to medium containing high concentration of cadmium. The selected cell lines, especially haploid cell line, were resistant an the high concentration of cadmium. And dry weight, ash weight, and cadmium contents of cell were increased. These results indicated that the selected cell lines showed higher resistance of cadmium than control cells, and haploid plant is more resistant than diploid plant on medium with cadmium.

담배(Nicotiana tabacum cv. BY4)의 이배체식물과 담배의 약으로부터 유도된 반수체식물의 잎절편을 사용하여 2.4-D를 단독톄처리하여 캘러스를 유도하였다. 캘러스증식은 이배체에서는 2.0 mg/L 2.4-D와 1.0mg/L 카이네틴의 조합처리구에서, 반수체의 경우에 있어서는 2.0mg/L 2.4-D와 0.1mg/L BAP 조합처리구에서 실시하였으며, 현탁배양도 동일조건으로 실시하였다. 현탁배양 세포괴를 배양 15-20일째가 되면 0.8% low melting agarose와 혼합하여 카드a뮴이 0-2,000 $\mu$M의 농도로 처리된 선발배지에 평판배양하여 카드뮴저항성 colony(cell clump)를 조사관찰하였다. 그 결과 본 실험에서는 이배체는 세포의 성장에 있어서 최하저해농도(MIC : Minimum Inhibition Concentration)는 Cd$^{++}$ 300 $\mu$M이었으며 반수체는 Cd$^{++}$ 200 $\mu$M이었다. 이때 반수체 및 이배체의 MIC에서 선발된 세포주를 카드뮴 500, 1,000, 2,000 $\mu$M 농도로 옳겨 경화시킨 후 카드뮴저항성을 선발하였고 그 특성을 조사하였다. 그 결과 카드뮴 저항성을 띤 반수체 및 이배체유래 캘러스의 경우에는 각각 카드a뮴의 농도가 증가할수록 생중량은 감소하였으나, 건조중량은 오히려 증가하였다. 그리고 각각 선발된 세포주내의 카드뮴의 함량을 AAS를 사용하여 측정하였던 바, 카드뮴의 농도가 높을수록 캘러스에 축적된 카드뮴의 양이 높게 나타났다.

Keywords

References

  1. Plant Cell Reports v.3 selection and characterization of cadmium-resistant suspension cultures of the wild tomato Lycopersicon peruvianum Bennetzen JL;Adams TL
  2. Plant Cell Line Selection. VCH Culture systems and selection procedures Dix PJ;Collin HA;PJ Dix(ed.)
  3. J Amer Soc Hort Sci v.113 Effects of simulated acid rain on cadmium mobilization in soils and subsequent uptake and accumulation in poplar and sunflower Gingas VM;Sydnor TD
  4. Plant Cell Reports v.9 Phytochelatin accumulation and stress tolerance in tomato cells exposed to cadmium Gupta SC;Goldsbrough PB
  5. Plant Cell Reports v.7 Cadmium tolerance in tabacco cell culture and its revance to temperature stress Huang SC;Goldsbrough PB
  6. Proc. Natl Acad Sci USA v.84 Poly(γ-glutamylycysteiny)glycines : Its role in cadmium resistance in plant cells Jackson PJ;Unkefer CJ;Doolen JA;Watt K;Robinson NJ
  7. In Vitro Cell Dev Biol v.25 Stress induced somatic embryogenesis in carrot and its application to synthetic seed production Kamada H;Kobayshi K;Kiyosue Harada
  8. Phytochemistry v.31 Phytochelaytins protect plant enzymes from heavy metal poisoning Kneer R;Zenk MH
  9. Korean J Limnology v.23 Cadmium detoxification mechanism of Cd-resistant Bacillus sp. isolated from industrisl sewage Lee KS;Kang SW;Kim YH;Kim EA;Kim KH;Choi YK
  10. Japan J Breed v.22 Competition among pollen grains for hapoid tobacco plant formation by anther culture Nakata K;Kurihara
  11. Korean J Plant Tissue culture v.21 Enzyme activity in plant regeneration from diploid and haploid calli of Nicotiana tabacum cv BY4 Oh SC;Soh WY;Cho DY;Yang DC
  12. Plant Physiol v.56 characterization of carrot and tobacco cell culture resistant to p-fluorophenylalanine Palmer JE;Widholm JM
  13. Plant Physiol v.98 Cadmium-sulfide crytallites in CD-(γEC)nG peptide complexes from tomato Reese RN;White CA;Winge DR
  14. M.S. thesis, Chunbuk National University Effects of cadmium on somatic embryogenesis and plant regeneration from cultures cells of Daucus L Shin EK
  15. Society for Experimental Biolory Seminar Series 49 : Inducible Plant Proteins Metal binding proteins and metal-regulated gene expression in higher plant Tomstt AB;Sewell AK;Jones SJ;Miranda JR;Thurman BA;J .L. Wary(ed.)
  16. Ann Rev Biochem v.41 Biochemical effects of mercury, casmium, and lead Vallee BL;Ulmer DD
  17. Plant Physiol v.92 Subcellular localization of cadmium and cadmium-binding prptides in tabacco leaves Vogelli-Large R;Wagner GJ