石蒜属植物遗传多样性的ISSR和RAPD标记比较研究

目的研究石蒜属4种植物的种间亲缘关系和同一种内不同采集地材料的遗传多样性,为其种质资源评价和合理开发利用提供分子佐证。方法用ISSR和RAPD标记分别对不同采集地的37份石蒜属植物(石蒜18份、中国石蒜10份、换锦花5份、忽地笑4份)的基因组DNA的遗传多样性进行检测。结果(1)16条ISSR和17条RAPD引物分别扩增出229和215条带,多态比率分别为85.59%和69.77%,显然,ISSR检测出的多态性条带的能力优于RAPD;(2)ISSR和RAPD标记检测同组供试材料种间或种内的Nei′s遗传相似系数范围分别为0.5459～0.9345,0.6419～1.0000,平均为0.6836,0.7428。两者相关系数r=0.8582,达极显著水平。(3)ISSR和RAPD标记的分子聚类结果相近,两种标记均能准确地把不同来源的材料先按种聚类,种间的分子分类结果与传统经典分类相符;种内不同采集地材料间存在一定的遗传差异,其遗传多样性较为丰富。(4)用POPGENE3.2分析的种间基因分化系数(Gst)与用AMOVA进行的遗传变异巢式方差分析所得结果基本一致。结论两种标记均可用于石蒜属植物种间亲缘关系与种内的遗传多样性研究,虽然ISSR标记在种间遗传多样性检测上分辨力较RAPD标记低,但ISSR标记能检测到更高的种内遗传差异性,更适合种下水平的遗传多样性研究。

Comparison between ISSR and RAPD markers in genetic diversity of plants in Lycoris Herb

Objective To investigate the inter-and intra-specific relationships among four species of Lycoris Herb.from different habitats and provide the evidence for the evaluation and exploitation of the germplasm of Lycoris Herb.Methods The genetic diversities of 37 accessions of Lycoris Herb.from different habitats were investigated with the technique of inter-simple sequence repeat(ISSR) and random amplified polymorphic DNA(RAPD),which were 18 of L.radiate,10 of L.chinensis,5 of L.sprengeri and 4 of L.aurea.Results(1) The 229 and 215 DNA bands were amplified with 16 ISSR and 17 RAPD primers respectively,the percentage of polymorphic bands(PPB) in ISSR detection(85.59%) was higher than that in RAPD(69.77%).(2) The coefficient ranges of interspecific and intraspecific genetic similarity(GS) in ISSR and RAPD analyses for the same materials were 0.545 9-0.934 5 and 0.641 9-1.000 0,respectively,the average were 0.683 6 and 0.742 8.There was significant difference of correlation coefficient at 0.01 level between ISSR and RAPD(r=0.858 2).(3) The similar molecular cluster results were obtained from RAPD and ISSR analyses,and showed that the genetic diversity of intraspecific was high,the interspecific relationship also matched well with the results of typical classification.(4) The coefficient of gene differentiation(Gst) among interspecific analyzed by POPGENE 3.2 consistent with the result of molecular variance analysis by AMOVA.Conclusion ISSR and RAPD markers were both efficient methods in revealing interspecific or intraspecific genetic differences and diversity in plants of Lycoris Herb.The acuity with ISSR detecting for interspecific genetic diversity is lower than that with RAPD.The more intraspecific genetic differences are dectected by ISSR marker.However,ISSR is more suitable for the study on intraspecific genetic diversity.