Welcome to Bulletin of Botanical Research! Today is Share:

Bulletin of Botanical Research ›› 2010, Vol. 30 ›› Issue (5): 588-593.doi: 10.7525/j.issn.1673-5102.2010.05.012

Previous Articles     Next Articles

Optimization of ISSR-PCR Reaction System and Primers Screening in Schisandra sphenanthera

LUO Cheng;XIONG Yu-Ting;GU Wei;*;WANG Zhe-Zhi;   

  1. 1.College of Life Science,Shaanxi Normal University,Xi’an 710062;2.Key Laboratory of the Ministry of Education for Medicinal Resources and Natural Pharmaceutical Chemistry,Xi’an 710062;3.National Engineering laboratory for Resource Development of Endangered Crude Drugs in Northwest of China,Shaanxi Normal University,Xi’an 710062
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-09-20 Published:2010-09-20
  • Contact: GU Wei
  • Supported by:

Abstract: The main influential factors of ISSR-PCR reaction system in Schisandra sphenanthera Rehd. et Wils. were systematically studied, optimization of ISSR-PCR system was established, and 12 effective ISSR primers were selected. Single-factor experimental results showed that the suitable concentration of composition of ISSR-PCR systen was 1.50~3.50 mmol·L-1 of Mg2+, 0.10~0.35 mmol·L-1 of dNTPs, 0.25~0.60 μmol·L-1 of primer and 0.50~1.50 U of Taq DNA polymerase, respectively. The optimal ISSR-PCR reaction system was defined by orthogonal experiment with four-factor of three-level, i.e. 20 μL reaction system contained 2.50 mmol·L-1 of Mg2+, 0.20 mmol·L-1 of dNTPs, 0.25 μmol·L-1 of primer, 1.50 U of Taq DNA polymerase, 60 ng of DNA template, 2.50 μL of 10×PCR Buffer. The present study could be used in the research for evaluation of germplasm resources and analysis of genetic diversity in S.sphenanthera Rehd. et Wils..

Key words: Schisandra sphenanthera Rehd. et Wils., ISSR, optimization of system, primers screening

CLC Number: