Welcome to Bulletin of Botanical Research! Today is Share:

Bulletin of Botanical Research ›› 2009, Vol. 29 ›› Issue (2): 234-241.doi: 10.7525/j.issn.1673-5102.2009.02.019

Previous Articles     Next Articles

Studies on Optimization of ISSR Amplified Conditions in Kudingcha Species in Oleaceae of China

ZHENG Dao-Jun;LIU Guo-Min;*;LIANG Yuan-Fa;YAN Dong-Hai;LINGHU Chang-Di;TIANG Yong-Hui   

  1. (1.Key Laboratory of Tropical Biological Researches. MOE,Hainan University,Haikou 570228) (2.Kudingcha Research Institute of Hainan University,Haikou 570228) (3.The Tea Research Institute of Guizhou Province,Meitan 564100)
  • Received:1900-01-01 Revised:1900-01-01 Online:2009-03-20 Published:2009-03-20
  • Contact: LIU Guo-Min
  • Supported by:

Abstract: The influential factors of ISSR for Kudingcha species in Oleaceae of China were systematically studied, a set of stable ISSR-PCR reaction parameters was established. 10 effective ISSR primers were selected out, and with them 21 test germplasm materials from 8 Kudingcha species in Oleaceae of China were examined for the repeatability and polymorphism of the optimized reaction conditions. The optimum ISSR-PCR reaction system was that 2.5 μL 10×PCR buffer, 2.0~3.0 mmol·L-1 MgCl2, 150~300 μmol·L-1 dNTPs, 1.0~1.5 Taq polymerase, 0.4~0.5 μmol·L-1 primers, and 5~320 ng DNA template were contained in 25 μL reaction solution. The optimized amplification program was that pre-denaturing at 94℃ for 4 min, then denaturing at 94℃ for 40 s, primer annealing at 50℃~54℃ for 45 s, extension at 72℃ for 120 s, for 35 cycles, at last extension at 72℃ for 8 min. The productions were stored at 4℃. The optimized ISSR-PCR reaction system was suitable for the study of genetic diversity and relationship of Kudingcha germplasm resources in Oleaceae of China.

Key words: Kudingcha species, Oleaceae, ISSR, influential factors, Optimization of system

CLC Number: