Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/70506
標題: Using ITS2 PCR-RFLP to generate molecular markers for authentication of Sophora flavescens Ait
作者: Lin, T.C.
Yeh, M.S.
Cheng, Y.M.
Lin, L.C.
Sung, J.M.
關鍵字: DNA marker
ITS2
PCR-RFLP
Sophora flavescens Ait
medicinal-plant
region
chromatography
期刊/報告no:: Journal of the Science of Food and Agriculture, Volume 92, Issue 4, Page(s) 892-898.
摘要: BACKGROUND: Dried root of Sophora flavescens Ait. is a medicinal material occasionally misused or adulterated by other species similar in appearance. In this study the internal transcribed spacer (ITS) regions of DNA samples of S. flavescens Ait. collected from different areas of Taiwan were amplified by polymerase chain reaction (PCR) and compared. The effectiveness of using ITS2 PCR restriction fragment length polymorphism (RFLP)-generated markers to differentiate S. flavescens Ait. from possible adulterants was also evaluated. BACKGROUND: Dried root of Sophora flavescens Ait. is amedicinal material occasionally misused or adulterated by other species similar in appearance. In this study the internal transcribed spacer (ITS) regions of DNA samples of S. flavescens Ait. collected from different areas of Taiwan were amplified by polymerase chain reaction (PCR) and compared. The effectiveness of using ITS2 PCR restriction fragment length polymorphism (RFLP)-generated markers to differentiate S. flavescens Ait. from possible adulterants was also evaluated. RESULTS: The S. flavescens Ait. samples collected from different areas were extremely low in ITS sequence variability at species level. ITS2 PCR-RFLP coupled with restriction enzymes Sac I, Sac II, Xho I or Pvu I produced specific fragments for all tested variants. ITS2 PCR-RFLP coupled with Sac II was further performed to identifymixtures of DNA extracts of S. flavescens Ait. and Sophora tomentosa L. in various ratios. The developed ITS2 PCR-RFLP markers could detectmixed DNA samples of S. flavescens Ait./S. tomentosa L. up to a ratio of 10 : 1. CONCLUSION: The present study demonstrates the usefulness of ITS2 PCR-RFLP coupled with pre-selected restriction enzymes for practical and accurate authentication of S. flavescens Ait. The technique is also suitable for analysing S. flavescens Ait. mixed with other adulterants. (C) 2011 Society of Chemical Industry
URI: http://hdl.handle.net/11455/70506
ISSN: 0022-5142
文章連結: http://dx.doi.org/10.1002/jsfa.4667
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