[关键词]
[摘要]
目的 获得在大青属Clerodendrum L. 主要树种间具有较好通用性的SSR标记,并构建DNA指纹图谱用于该属重要民族药用植物近缘种的分子甄别和遗传学研究。方法 选用2种大青属植物伊豆大青C. izuinsulare和海州常山C. trichotomum的19对SSR引物,对大青属9个种共9个样本进行扩增检测。结果 17对引物在广布种大青中有扩增产物,通用率最高(89.5%),另有7对引物在所有样本中有扩增产物,通用率为36.8%。采用多态性较好的6对SSR引物构建9个种的DNA指纹图谱,其中引物CI140可鉴定除白花灯笼和尖齿臭茉莉之外的7个种,该引物与CI107、CI132、CI143和CT042两两组合可鉴别所有9个种;利用遗传距离进行UPGMA聚类,结果显示腋序系白花灯笼单独成一组,海通和大青成一组,其他聚类分组结果与其形态分类存在一定差别。结论 近缘种间共用SSR引物是开发大青属植物SSR标记行之有效的方法,可为大青属植物分子标记资源的使用提供参考。
[Key word]
[Abstract]
Objective In order to obtain SSR primers which has good universality among the main tree species of Clerodendrum L., and then molecular screening of DNA fingerprints was used in closely related species and genetic research of these ethnical medicinal resources. Methods An experiment was conducted to study the transferability of 19 pairs of SSR primers developed from C. izuinsulare and C. trichotomum and amplified in nine samples from nine species of Clerodendrum L. Results Seventeen pairs of 19 primers had the amplification products in C. cyrtophyllum, a cosmopolitan species, the transferability ratio was the highest (89.5%). Seven pairs of primers were successfully amplified in all samples of Clerodendrum L. species, and the transferability ratio was 36.8%. Six pairs of polymorphism SSR primers were used to construct the DNA fingerprints of nine Clerodendrum L. species. In the primers, the CI140 identified seven Clerodendrum L. species except C. fortunatum and C. lindleyi. Coupled with CI132, CT042, CI107, or CI143, they identified all of the nine Clerodendrum L. species. Then, the genetic distances were used to generate a UPGMA tree. The results showed that C. fortunatum, belonging to Ser. Axilliflorae schauer, formed a cluster, and C. mandarinorum and C. cyrtophyllum formed another cluster, while the clusters of the others were difference from the shape classifications. Conclusion The experiment proves a good way to develop Clerodendrum L. SSR primers from other related species. This study provides the important reference for the use of molecular markers of Clerodendrum L.
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[基金项目]
江西省科技支撑项目(20142BBF60031);江西省林业科学院青年培养项目(2011521201)