摘要： 利用公共数据库已有的ISSR序列信息，检验了29条ISSR引物在谷子等狗尾草属植物基因组的扩增性，其中14条在狗尾草属中有良好扩增，其余1条在狗尾草属中无任何扩增。能稳定扩增的14条引物全为3'-锚定引物，6条'-锚定引物在狗尾草属中得不到扩增。研究发现AG，GA和CA重复的引物在狗尾草属植物有较好扩增，且多态性强。利用正交试验设计，从Mg²⁺、dNTP、引物、Taq酶4种因素、4个水平对狗尾草属谷子ISSR反应体系进行优化，确定了适合谷子的ISSR反应体系，在20 μL体系中，含1×PCR buffer、Mg²⁺浓度2.0 mmol/L，dNTP浓度0.2 mmol/L，Taq酶0.4 μL，引物浓度0.2 μmol/L，0 ng模板，通过梯度PCR确定了引物的退火温度。本研究筛选的ISSR引物及建立的优化反应体系可用于谷子等狗尾草属植物遗传多样性和基因组研究，并对其他近缘作物有指导意义。

关键词: 谷子, ISSR标记, 狗尾草属

Abstract: Twenty nine ISSR primers designed according to the public ISSR database were tested their reaction feasibility in foxtail millet and other Setaria species genomes, of which 14 primers gave perfect amplication profile and the other 15 made no amplication. All the 14 primers that made good amplication were 3'-anchored. The results also showed that poly (AG) or (GA) and poly (CA) have better amplictiaon and polymorphism than other primers in Setaria genomes. The orthogonal design was used to optimize ISSR amplification system of Setaria in four factors (Mg²⁺, dNTP, primer, Taq polymerase) at four levels respectively. A suitable ISSR reaction system was established, namely 20 μL reaction system containing 50 ng DNA templet, 0.25 μmol/L primer, 1×PCR buffer, 0.25 mmol/L dNTP, 0.4 μL Taq DNA polymerase, 2.0 mmol/L Mg²⁺. The results obtained can be used for Setaria genetics and genome research, and are important for related sepecies in connected fields.

Key words: Foxtail millet, ISSR marker, Sataria

中图分类号: 

• S515.01