安徽农业科学2011，Vol.39Issue(27)：16490-16493,4.
真藓科植物ISSR-PCR反应体系的优化及ISSR指纹图谱的初步构建
Optimization of ISSR-PCR Reaction System and Preliminary Construction of ISSR Fingerprinting of Some Species in Bryaceae
摘要
Abstract
[Objective] The aim was to provide molecular basis for the identification of species in the moss family Bryaceae by the construction of inter simple sequence repeats (ISSR) fingerprinting. [ Method] In order to seek standardizing PCR reaction set-up, orthogonal design was used to optimize 1SSR-PCR amplification system of Bryaceae in five factors (Mg2+ , dNTPs, primer, DNA template, Taq DNA polymerase) at four levels respectively. [Result] A suitable ISSR reaction system was obtained, namely: 20 μl reaction system containing 5 ng DNA template, 0.2μmol/L primer, 2.25 mmol/I. MgCl2, 0. 6 U Taq DNA polymerase, 0.4 mmol/I. dNTPs. Proper annealing temperature was 48 - 50 ℃. The above system and six ISSR-PCR primers were used for the PCR amplification of 14 samples from Bryaceae and the related species in Mniaceae. A total of 86 bands were amplified, of which 86 (100% ) was polymorphic. NJ cluster analysis showed a stellate cladogram. [Conclusion] The results manifested that ISSR fingerprinting could provide the appropriate degree of polymorphism at low taxonomic level, so it would be a useful tool to provide additional evidence for resolving taxonomic relationships at the species level of Bryaceae.关键词
真藓科/ISSR/反应体系优化/种级水平/分类关系Key words
Bryaceae/ ISSR/ Optimization of reaction system/ Species-level/ Taxonomic relationship分类
农业科技引用本文复制引用
汪琛颖,赵建成..真藓科植物ISSR-PCR反应体系的优化及ISSR指纹图谱的初步构建[J].安徽农业科学,2011,39(27):16490-16493,4.基金项目
河北省自然科学基金资助项目(C2006000147) （C2006000147）
郑州市科技计划资助项目(10PTGN449-6) （10PTGN449-6）
河南省科技计划项目(112300410018). （112300410018）
