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:25μL反应体系中DNA模板的量为30.00ng、引物0.60μmol/L、dNTP 0.20mmol/L、Taq酶0.75U;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。
Taking Cypri pedium macranthurn , Cypri pediurn calceolus ,Cypri pedium ventricosum ,Cypri pedium 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.
出处
《北方园艺》
CAS
北大核心
2013年第24期92-95,共4页
Northern Horticulture
基金
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)
