Journal of South China University of Technology (Natural Science Edition) ›› 2009, Vol. 37 ›› Issue (6): 74-78,83.

• Biological Engineering • Previous Articles     Next Articles

Molecular Identification of Fallopia multiflora by PCR-RFLP Based on rDNA ITS Sequence

Zheng Chuan-jin1  Zhao Shu-jin2  Zhao Zhen-hua1  Guo Jun1   

  1. 1 School of Biological Science and Engineering, South China University of Technology, Guangzhou 510006, Guangdong, China; 2. General Hospital of Guangzhou Military Command, Guangzhou 510010, Guangdong, China
  • Received:2008-06-05 Revised:2008-07-31 Online:2009-06-25 Published:2009-06-25
  • Contact: 赵树进(1958-),男,教授,博士生导师,主要从事生物制药研究.E-mail:gzzsjzhs@163.com E-mail:z_c_jin@163.com
  • About author:郑传进(1973-),男,博士生,主要从事药用植物分子鉴别研究.
  • Supported by:

    广东省自然科学基金资助项目(6104397)

Abstract:

The rDNA internal transcribed spacers (ITS) of Fallopia multiflora (F. multiflora) and its frequent adulterants, such as Fallopia multiflora var. cilliinerve, Pteroxygonum giraldiiwere and Cynanchum auriculatum, were sequenced and analyzed. The results show that in the ITS1 region, the divergences between F. multiflora and the three adulterants are respectively 6.91%, 18.10% and 42. 20%, while in the ITS2 region, the divergences are respectively 10. 00%, 19.40% and 43.90% ; and that the intraspecies ITS1 and ITS2 divergences of F. multi- flora are 0. 00% -2. 13% and 0. 00% -1.03% , respectively. Based on the divergences, a endonuclease Nsb I restriction site specific for F. multiflora was detected in the ITS2 region and was used for PCR-RFLP (Polymerase Chain Reaction and Restriction Fragment-Length Polymorphism) analysis. It is found that the PCR product from F. multiflora rDNA ITS can be cleaved by Nsb I into two fragments of about 531 bp and 109bp, while that from the adulterants cannot be cleaved, and that the PCR-RFLP map still retains the undigested original rDNA ITS pattern. It is thus concluded that the proposed PCR-RFLP method is effective in discriminating F. multiflora from its adulterants.

Key words: Fallopia multiflora, molecular identification, rDNA internal transcribed spacer, PCR-RFLP