SUN Ye-ying,CHEN Li-fei,LIU Shu-ying,et al.Establishment of the Optimum ISSR-PCR Reaction System of Cypripedium in the Region of Changbai Mountain of China[J].Northern Horticulture,2013,37(24):92-95.
长白山区杓兰属植物ISSR-PCR最佳反应体系的建立
- Title:
- Establishment of the Optimum ISSR-PCR Reaction System of Cypripedium in the Region of Changbai Mountain of China
- 文章编号:
- 1001-0009（2013）24-0092-04
- Keywords:
- Changbai mountain; Cypripedium; ISSR; reaction system
- 分类号:
- S 682.31
- 文献标志码:
- A
- 摘要:
- 以长白山区的5种杓兰属植物大花杓兰、杓兰、东北杓兰、山西杓兰、斑花杓兰为试材，对影响PCR扩增效果的模板浓度、引物浓度、dNTP浓度、Taq酶量以及退火温度进行了优化，以期建立杓兰属植物ISSR-PCR的反应体系。结果表明：25 μL反应体系中DNA模板的量为30.00 ng、引物0.60 μmol/L、dNTP 0.20 mmol/L、Taq酶0.75 U；该体系以5种杓兰属植物，4个引物进行扩增验证，能够得到清晰的条带，且具有多态性，而且有可重复性；表明建立的杓兰属ISSR-PCR体系适用遗传多样性的研究。
- Abstract:
- Taking Cypripedium macranthum，Cypripedium calceolus，Cypripedium ventricosum，Cypripedium shanxiense and Cypripedium guttatum of 5 Cypripedium species which distributed in Changbai mountains as test materials，the 4 factors and annealing temperatures which affect the amplification of PCR were optimized.The optimal ISSR-PCR conditions for Cypripedium were established.The results showed that containing of 30.00 ng template DNA，0.60 μmol/L primer，0.20 mmol/L dNTP，and 0.75 U Taq DNA polymerase in a volume of 25 μL was optimal.The clear，pleomorphic and repetitive amplified bands might be obtained with the above ISSR-PCR reaction conditions by 5 Cypripedium and 4 primers.It was revealed that the ISSR-PCR reaction system was suitable for researching of genetic diversity.
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备注/Memo
第一作者简介：孙叶迎（1989-），女，硕士研究生,研究方向为分子标记。Email:sunyeying3928@163.com.
责任作者：刘洪章（1957-），男，博士，教授，博士生导师,研究方向为植物资源。Email:lhz999@126.com.
基金项目：农业部野生植物资源调查资助项目;吉林省科技厅资助项目（20100254）。
收稿日期：2013-09-13