矮牡丹与紫斑牡丹RAPD分析初报

PRELIMINARY REPORT OF RAPD ANALYSIS IN PAEONIA SUFFRUTICOSA SUBSP.SPONTANEA AND PAEONIA ROCKII

用10个任意序列的寡核苷酸片段作为引物,将采自陕西、山西、甘肃等地矮牡丹与紫斑牡丹基因 组DNA,在毛细管气浴式PCR热循环仪上随机扩增。在对所有引物扩增条件严格标准化的条件下,这些引物可产生清楚的、可重复的与扩增产物相应的琼脂糖凝胶电泳区带。这10个有效扩增的引物在矮牡丹平均每一个体中扩增出71条带,其中多态的带16条(22.5％);在紫斑牡丹平均每一个体中扩增出76条带,其中多态的带21条(27.6％)。对每两个个体,每条多态带进行成对比较,累加后求分子标记差异的平均值。在矮牡丹3个居群间的平均差异是7.9;在紫斑牡丹4个居群间的平均差异是8.7;在矮牡丹与紫斑牡丹2个种间的平均差异是10.3。显然,若用来扩增的植物个体数目和引物数目增加,将会得到更满意的结果。初步结果表明濒危植物矮牡丹与紫斑牡丹种内低水平的DNA多态性。RAPD技术用于检测野生牡丹居群内与居群间的遗传变异是有用的与可行的;用于研究种间的进化和亲缘关系也有潜力。

Ten different oligonucleotide primers of arbitrary sequence were used in Capillary and Air Thermocycler to amplify genomic total DNA of Paeonia suffruticosa subsp. spon-tanea and P. rockii isolated from several local populations in Shanxi, Shaanxi and Gansu provinces. Under the strictly standardized amplification condition for all the primers, these primers yielded clear and reproducible bands corresponding to amplified products and separable by agarose gel electrophoresis. Among a total of 71 bands amplified, 16(22. 5%) were polymorphic in a single individual of P. suffruticosa subsp. spontanea, while among a total of 76 bands 21(27. 6%)were polymophic in a single individual of P. rockii. On an average, the pairwise marker difference between band profiles of conspecific individuals (different populations) was 7. 9 for P. suffruticosa subsp. spontanea and 8. 7 for P. rockii respectively. The average marker difference between P. suffruticosa subsp. spontanea and P. rockii was 10. 3. Obviously, greater number of plants and primers will be required to detect satisfactorily level of genetic diversity. These preliminary results showed that intraspecific genetic diversity was low for the two endangered species. RAPD as a molecular marker was useful and feasible for detecting the genetic variation within species of wild moutans. And it was also potential for studying evolution and relationships between species.