红皮云杉与嫩江云杉RAPD和ISSR分子标记反应体系优化和特异性检测

以红皮云杉和嫩江云杉为材料，采用现代分子标记技术-RAPD和ISSR标记研究红皮云杉与嫩江云杉间在基因组水平上的遗传差异。通过正交试验设计筛选RAPD和ISSR反应程序和反应体系。从近100个引物中筛选出3个RAPD引物和一个ISSR引物用来区分红皮云杉和嫩江云杉，它们分别是OPN07、OPA17、S25和ISSR67。用这4个引物扩增两种云杉基因组DNA，分别在1 000，950，1 500，2 000 bp处嫩江云杉有特异性谱带而红皮云杉没有。研究表明，采用RAPD和ISSR分子标记可以将红皮云杉和嫩江云杉在分子水平上加以区分，为今后进一步阐明云杉的系统进化，物种鉴定和种间杂交育种提供了理论依据。

Reaction System Optimization and Specific Identification of Picea koraiensis and P.koraiensis var. nenjiangensis by RAPD and ISSR Marker

Picea koraiensis and Picea koraiensis var. nenjiangensis were used as the materials. The genetic differences in genomic level between Picea koraiensis and its variety were identified using RAPD and ISSR markers. The optimal reaction system and program of the markers were found through the orthogonal experimental design. Three RAPD primers OPN07，OPA17，S25 and one ISSR primer ISSR67 were screened out to make identification between Picea koraiensis and Picea koraiensis var. nenjiangensis, in which the specific bands were amplified in genomic DNA of Picea koraiensis var. nenjiangensis at 1 000, 950, 1 500 and 2 000 bp in molecular size, but not in Picea koraiensis. The results indicated that RAPD and ISSR marker could be used to differentiate these two spruces. In conclusion, two spruce species were identified in molecular level in this study. The results will provide basic information for systematical evolution, species identification, and interspecies crossing of spruce in the future.