Selection of Tropane Alkaloids High-Producing Lines by Single Cell Cloning of Hyoscyamus niger L. Root Cultures

  • Min, Ji-Yun (Division of Environmental Forest Science, Gyeongsang National University(Institute of Agriculture and Life Science, Gyeongsang National University)) ;
  • Park, Dong-Jin (Division of Environmental Forest Science, Gyeongsang National University(Institute of Agriculture and Life Science, Gyeongsang National University)) ;
  • Jeong, Mi-Jin (Division of Environmental Forest Science, Gyeongsang National University(Institute of Agriculture and Life Science, Gyeongsang National University)) ;
  • Song, Hyun-Jin (Division of Environmental Forest Science, Gyeongsang National University(Institute of Agriculture and Life Science, Gyeongsang National University)) ;
  • Kang, Seung-Mi (Division of Forest Research, Gyeongsangnam-do Forest Environment Research Institute) ;
  • Kang, Young-Min (College of Forest Resources, Forest and Wildlife Reserch Center, Department of Forest Products, Mississippi State University) ;
  • Choi, Myung-Suk (Division of Environmental Forest Science, Gyeongsang National University(Institute of Agriculture and Life Science, Gyeongsang National University))
  • Received : 2008.11.10
  • Accepted : 2009.03.04
  • Published : 2009.04.30

Abstract

Hyoscyamus species is sources of the hypnotic and sedative drugs hyoscyamine and scopolamine. Single cells of Hyoscyamus niger were dissociated from suspension cultures and adventitious roots obtained from single-cell clones which were cultured on B5 medium containing 3% (w/v) sucrose, 0.1 mg/L IBA and 0.4% (w/v) gelrite. H. niger adventitious root lines showed wide variation in tropane alkaloids production and growth. An effective selection of 200 root lines was made possible by the application of the 'Dragendorff's reagent' for qualitative detection of the alkaloids from root. A high correlation coefficient (r=0.9390) was observed between the values obtained with the two methods based on HPLC and Dragendorff's reagent analysis. Among the selected roots, the highest scopolamine content was 16.64 mg/g DW (Hn-59), which was 8.82-fold more productive than the lowest alkaloid producing line (Hn-25). Here, we established an efficient selection method on tropane alkaloids production and suggest that the Dragendorff's reagent is of great practical value in selection of invisible compounds.

Keywords

Acknowledgement

Supported by : Korea Research Foundation

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