Using ITS2 PCR-RFLP to generate molecular markers for authentication of Sophora flavescens Ait.
Version of Record online: 7 OCT 2011
Copyright © 2011 Society of Chemical Industry
Journal of the Science of Food and Agriculture
Volume 92, Issue 4, pages 892–898, 15 March 2012
How to Cite
Lin, T. C., Yeh, M. S., Cheng, Y. M., Lin, L. C. and Sung, J. M. (2012), Using ITS2 PCR-RFLP to generate molecular markers for authentication of Sophora flavescens Ait. J. Sci. Food Agric., 92: 892–898. doi: 10.1002/jsfa.4667
- Issue online: 6 FEB 2012
- Version of Record online: 7 OCT 2011
- Manuscript Accepted: 15 AUG 2011
- Manuscript Revised: 10 JUL 2011
- Manuscript Received: 16 FEB 2011
- DNA marker;
- Sophora flavescens Ait.
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.
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 identify mixtures of DNA extracts of S. flavescens Ait. and Sophora tomentosa L. in various ratios. The developed ITS2 PCR-RFLP markers could detect mixed 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. Copyright © 2011 Society of Chemical Industry