药用植物厚朴TRAP标记的开发和利用

    Development and Utilization of TRAP Molecular Markers on Medicinal Plant Magnolia officinalis

    • 摘要:
      目的 开发厚朴新型分子标记,分析广东乐昌龙山林场厚朴资源的亲缘关系,以方便后续进行厚朴优株选择和优良品种选育等研究工作。
      方法 以14个由广东乐昌龙山林场种植的、种源来自江西省各地的药用植物厚朴为材料,根据GenBank中厚朴cDNA序列信息,利用NCBI(https://www.ncbi.nlm.nih.gov)上Primer BLAST工具设计厚朴靶区域扩增多态性(Target Region Amplified Polymorphism, TRAP)标记固定引物,与SRAP(Sequence Related Amplified Polymorphism)随机引物配对,扩增厚朴基因组DNA,开发厚朴TRAP标记。统计TRAP标记与厚朴SSR(Simple Sequence Repeat)标记扩增产物的电泳带型图谱,用NTSYSpc2.10e软件进行聚类分析,分析14个厚朴资源的遗传多样性及其亲缘关系。
      结果 从14条固定引物与5条随机引物配组形成的70对引物中共开发出7个扩增产物电泳条带清晰、带型丰富、多态性好的厚朴TRAP标记;构建了厚朴资源的亲缘关系树状图,结果表明龙山林场厚朴资源遗传相似系数在0.75~0.88之间,母株与对应子代之间遗传相似系数并不是最高。
      结论 厚朴群体内基因交流频繁,遗传多样性丰富,新开发的TRAP标记将为厚朴优株鉴别、优良品种选育、分子标记辅助选择等后续研究工作提供便利。

       

      Abstract:
      Objective The study was carried out to develop new molecular markers and analyze the molecular phylogeny of Magnolia officinalis in Longshan Tree Farm, Lechang, Guangdong, with a view to facilitating subsequent researches on the selection of excellent plants and breeding of superior varieties.
      Method Fourteen Magnolia officinalis (provenances from Jiangxi Province) individual plants collected from Longshan Tree Farm were used as tested materials. By using Primer BLAST on NCBI (https://www.ncbi.nlm.nih.gov), fixed primers of TRAP molecular markers were designed based on the sequences of M. officinalis cDNA in the GenBank. The fixed primers were matched with random primers (sequence related amplified polymorphism) to amplify genomic DNA and develop TRAP molecular markers. The electrophoretogram of TRAP markers and amplified products of M. officinalis SSR (Simple Sequence Repeat) were counted. Then, NTSYSpc2.10e was used for cluster analysis to discover the genetic diversity and phylogeny of fourteen M. officinalis resources.
      Result Seven TRAP molecular markers showing good polymorphism and clear electrophoresis bands were successfully developed from seventy primer pairs formed by fourteen fixed primers and five random primers. The phylogenetic tree of M. officinalis plants was constructed. The genetic similarity coefficient of these M.officinalis plants was 0.75-0.88 and it was not the highest between mother plant and its offspring.
      Conclusion M. officinalis was rich in genetic diversity and gene exchange was frequent in M. officinalis population. The newly developed TRAP molecular markers will facilitate identification of superior plants, breeding of excellent varieties and marker-assisted selection of M. officinalis in the future.

       

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