海南龙血树ISSR-PCR反应体系建立与有效引物筛选  

Establishment of ISSR-PCR Reaction System and Screening of Effective Primers with ISSR Marker for Dracaena cambodiana

作　　者：郑道君[1] 谢良商[1] 曾建华[1] 张治礼[1] 

机构地区：[1]海南省农业科学院农作物遗传育种重点实验室,海口571100

出　　处：《热带亚热带植物学报》2011年第2期177-183,共7页Journal of Tropical and Subtropical Botany

基　　金：国家自然科学基金项目(30760015);海南省自然科学基金项目(30831)资助

摘　　要：采用单因子试验与正交试验的方法,研究了海南龙血树(Dracaena cam bodiana Pierre ex Gagnep)ISSR-PCR反应体系中的主要影响因子,筛选出16条有效引物并优化了它们的退火温度,此外还检测了体系的重复性。优化后的25μL反应体系包含:10×PCR buffer2.5μL,3.0mmol/LMgCl2,300μmol/LdNTPs,Taq聚合酶3.0U,引物0.2μmol/L,DNA模板20ng。该反应体系可用于海南龙血树的遗传多样性和遗传结构的分析。The influence factors of ISSR PCR for Dracaena cambodiana Pierre ex Gagnep were studied by using single factor test and orthogonal design, and a set of ISSR PCR reaction system was established. Sixteen ISSR primers were selected out, and their annealing temperatures were also optimized. Furthermore, the optimized reaction conditions were examined for its repeatability in D. cambodiana. The optimum ISSR PCR reaction system with total 25 μL reaction solution contain 2.5 μL 10 x PCR buffer, 3.0 mmol/L MgCl2, 300 Izmol/L dNTPs, 3.0 U Taq polymerase, 0.2 μmol/L primers, and 20 ng DNA template, which can be used for analysis in genetic diversity and genetic structure of D. cambodianas.

关 键 词：海南龙血树 ISSR 影响因子 体系优化 引物筛选 

分 类 号：S567.19[农业科学—中草药栽培]