摘要 | 以陕西产野生石蒜〔Lycoris radiata (L’Hr.)Herb.〕为材料,通过单因子实验分别研究了模板DNA、Mg2+、dNTPs、引物浓度及Taq DNA聚合酶用量对石蒜SRAP分子标记扩增体系的影响,并对扩增体系进行了优化。优化后的反应体系总体积为10μL,含20ng模板DNA、3.0mmol·L-1Mg2+、0.20mmol·L-1dNTPs、0.4μmol·L-1引物和0.50UTaq DNA聚合酶。运用优化体系对9个石蒜居群的基因组DNA进行扩增,获得的DNA条带清晰,多态性比较丰富。说明SRAP-PCR可用于石蒜属植物的亲缘关系、系统演化、物种鉴别和遗传多样性等领域的研究。 |
Abstract | Effects of concentrations of template DNA, Mg2+, dNTPs, primers and amount of TaqDNA polymerase on SRAP molecular marker amplification system of Lycoris radiata(L' Hér.)Herb. from Shaanxi Province were studied by monofactorial experiment, and amplification system was optimized, too. Total volume of optimal system is 10μL, which contains 20ng template DNA, 3.0 mmol·L- 1 Mg2+,0.20mmol· L- 1dNTPs, 0.4μmol· L- 1primerand 0.50U TaqDNA polymerase. By using this optimal system, genomic DNA of nine populations of L. radiate was amplified, and DNA bands were clear and had high polymorphism. It is concluded that SRAP-PCR can be used to research genetic relationship, systematic evolution, species identification and genetic diversity of Lycoris Herb. species. |
关键词 | 石蒜; SRAP-PCR; 扩增体系优化; 遗传多样性; |
Key words | Lycoris radiata(L' Hér.)Herb.; SRAP-PCR; optimization of amplification system; genetic diversity |
作者 | 袁菊红1,2,权俊萍1,胡绵好3,孙 视1,彭 峰1,夏 冰1 |
所在单位 | 1.江苏省•中国科学院植物研究所(南京中山植物园),江苏南京210014; 2.江西财经大学资源与环境管理学院,江西南昌330032; 3.上海交通大学农业与生物学院,上海201101 |
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基金项目 | 江苏省道地药材种质资源库建设项目(BM2006104); 江苏省植物迁地保护重点实验室开放基金资助项目(KF2007001); |