【作者】 刘军；

【导师】 刘学端；

【作者基本信息】 中南大学 ， 生物工程， 2012， 硕士

【摘要】 药用植物基因多样性即遗传多样性,是指蕴藏在植物个体基因里的所有遗传信息的总和,包括一个群体内不同个体或种内不同群体之间的遗传变异。全面准确地了解药用植物基因多样性,将能探明其进化的历史,有助于资源开发、物种保护及种资源保护。传统中药多是复方制剂,成分多样且作用机制复杂,这给药物研究带来很大的困难。而基因芯片技术具有大规模、高通量、平行检测等优势,是从基因层面来鉴别植物,不会受植物不同取样部位、不同取样时间以及不同前处理过程与方式的影响,结果稳定,重复性好。重楼属植物是极具药用价值的植物,但长期以来其野生资源被过度采集,在许多地区已濒临灭绝,因此开展其物种分子水平的研究很有必要。本研究测定了壶瓶山8种重楼属样品rbcL基因、atpB基因和18SrDNA基因序列,比较了该3个基因的差异,构建了基于这3个基因序列的遗传进化树,分析它们的同源性关系,对其属下分类及亲缘关系进行了描述,探讨重楼属植物的基因多样性。利用现有药用植物基因资源筛选其特异性片段制作探针。然后用接触式点样法构建用于药用植物分析的基因芯片,优化了芯片杂交的关键技术,探索最优杂交温度为55℃。用基因芯片检测了10种药用植物：长春花、银杏、青蒿、文县黄耆、杜仲、黄连、野薄荷、红豆杉、华中雪莲、黄芩。长春花的功能基因最丰富,进而富含大量的次生代谢产物如黄酮、生物碱、萜类等。最后将植物群落基因组芯片鉴别了凌云重楼和壶瓶山重楼,结果表明凌云重楼比壶瓶山重楼功能基因丰富,有效成分含量更高,更具有药用价值。更多还原

【Abstract】 Genetic diversity of medicinal plant means heredity diversity, that is, the sum of all heredity information which is embodied in individual genes of plants, including genetic variation from different individuals within a group or different groups within a species. A comprehensive accurate understanding of the genetic diversity of medicinal plants will explore the evolutionary history of them and be helpful to resource development, species protection and the protection of seed resources.Traditional Chinese medicine has always been compound preparation, diversified in the composition and complex for the mechanism action, which caused great difficulties for drug research. The gene chip technology has the advantage of large scale, high throughput and parallel detection as an effective screening method. Gene chips identify plants from genetic level, not influenced by different sampling parts of the plant, different sampling time, pre-process and the ways. And the results are stable and good in repeatability.The Paris L is of great medicinal value and is one of the main raw materials of a variety of Chinese patent medicine. But for a long time the wild resources have been collected excessively, and even in many areas they are on the verge of extinction. For the protection of species resources and the genetic diversity, it becomes quite necessary to research on the molecular level of the species. This study measured rbcL gene, atpB gene and18SrDNA gene sequences of8Paris L samples, comparing their difference, forming the genetic evolutionary tree based on the3gene sequences, analyzing their homology relationship, describing the subordinate classification and genetic relationship, and discussing the genetic diversity of the Paris LThis paper states how to produce probe choosing their specific fragments by screening of the existing medicinal plant genetic resources and how to construct gene chips for the analysis of medicinal plants through contact styled spotting, optimising the critical technology of chip hybridization and discover the most favorable temperature,55℃, for the optimal hybridization.10medicinal plants have been detected using gene chips:Catharanthus roseus, Ginkgo biloba, Artemisia apiacea, Astragalus wenxianensis, Eucommia ulmoides, Coptis chinensis, Mentha haplocalyx, Taxus chinensis, Saussurea veitchiana Drumm, Scutellaria baicalensis. The results showing Catharanthus roseu is most abundant in functional genes, and it contains large amounts of secondary metabolites such as flavonoids, alkaloids and terpenes. At last, Paris cronquistii and Paris hupingshanensis have been identified through plant community genomic microarrays, the result of which shows that Paris cronquistiiha is more abundant in functional genes, active ingredients and medicinal value than Paris hupingshanensisis.更多还原