诺丽叶片DNA的提取及ISSR-PCR反应体系的建立

Isolation of Genomic DNA in Morinda citrifolia Linn.and Establishment of Its ISSR-PCR Reaction System

[目的]提取诺丽(Morinda citrifoliaLinn.)叶片DNA,并建立ISSR-PCR反应体系。[方法]以3份采自海南、4份采自美国的诺丽种质的叶片为材料,对其DNA提取和ISSR分子标记方法进行了研究。[结果]采用改进的CTAB DNA微量提取法,可以得到高质量的诺丽叶片基因组DNA。用14条不同的ISSR引物对所提取的诺丽基因组DNA进行了ISSR分子标记分析,其中7条引物在诺丽DNA中可扩增出多态性产物。[结论]建立了诺丽叶片基因组DNA快速、高效的提取方法和ISSR标记体系,可为ISSR分析应用于诺丽遗传研究奠定良好的基础。

[Objective] The research aimed to isolate the genomic DNA in Morinda citrifolia Linn.and establish its ISSR-PCR reaction system.[Method] The Morinda citrifolia Linn.leaves of 3 germplasm resources from Hainan and 4 germplasm resources from America were chose as materials,the methods of isolating DNA in Morinda citrifolia Linn.and establishment of its ISSR-PCR were studied.[Result]The results showed that the good quality DNA of Morinda citrifolia Linn.leaves were gained by the modified CTAB method.The isolated DNA could be used for ISSR analysis and 7 of 14 ISSR primers revealed polymorphisms in the Morinda citrifolia Linn.leaves DNA.[Conclusion]The rapid and efficient extraction method and the ISSR analysis procedure in this experiment laid a good foundation for application of ISSR in Morinda citrifolia Linn.genetic research.