Optimization of ISSR-PCR Reaction System and Preliminary Construction of ISSR Fingerprinting of Some Species in Bryaceae

[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, an orthogonal design was used to optimize ISSR-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 of DNA template, 0.2 μmol/L primer, 2.25 mmol/L MgCl2, 0.6 U of Taq DNA polymerase, 0.4 mmol/L dNTPs. Proper annealing temperature was found at 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, all showed polymorphism. NJ cluster analysis showed a star-shaped 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.

Study on Establishment of cpSSR Marker Technology and Optimization of Its Reaction System in Jatropha curcas Linn

[ Objective] The aim of this study was to establish the optimum cpSSR-PCR system for Jatropha curcas Linn. [ Method] cpSSR-PCR amplification system for Jatropha curcas Linn influenced by five factors including Taq DNA polymerase, Mg^2＋ , DNA template, dNTP and primer were optimized from several levels. [ Result] The optimum concentration of 20 μl reaction system was 10 × Buffer, 2.00 mmol/L Mg^2＋ , 2 U/μl Taq DNA polymerase, 0.2 mmol/L dNTP, 0.2 μmol/L primer and 35 ng/μl DNA template. [ Conclusion] The optimum annealing temperature for cpSSR-PCR reaction system is 52 ℃, and the cpSSR reaction system is steady and reproducible.

Controlled Synthesis of Polymers

Controlled synthesis is central to obtaining polymers with accurate structures and excellent performances.Recent research in the controlled synthesis of polymers has focused on optimizing monomers,initiation systems,and reaction conditions.The satisfactory sequence,topological structure,and dispersity have been achieved to satisfy the growing demand for functional polymers.This re-view summarizes the selection of monomers of various types and structures,the innovation of initiation systems,and the optimiza-tion of reaction conditions in the controlled synthesis of polymers and discusses their challenges and opportunities.