鸦片罂粟的基因组学研究及高特异性遗传分子标记筛选

As the raw plant for opium and heroin, poppy cultivation has been regulated strictly. Until now, the studies on poppy for highly specific markers screening, DNA polymorphisms, genetic diversity, high-density linkage map constructing, and phylogenic relationship are rare. This study uses comparative genomics and sequencing technology, trying to do the research of opium poppy from system evolution, gene family clustering, important medicinal ingredient synthesis pathway, fruit development molecular pathway, construction of genetic mapping and discovery of highly specific markers and other aspects. The completion of whole genome sequencing of Papaver somniferum would provide strong theoretical basis of parsing the time that core eudicots differentiated from Ranunculales. It would be significant for exploring the origination and evolution of eudicots basal groups, clarifying the relationship between whole genome duplication of Papaveraceae and the differentiation of closely related species, answering the evolutionary position of poppy in Ranunculales from genomic level and evolutionary mechanisms of genome size of P.somniferum. Moreover, opium poppy are widely distributed and diverse varieties, after obtaining genomic information, we can develop appropriate molecular markers for different varieties. This research could provide a theoretical basis for the identification of opium poppy and identification the origin of the drugs. In addition, by revealing the metabolic pathway of important medicinal components and mechanisms of regulatory genes for oil producing, this research would provide molecular basis for increasing medicinal component and oil content of poppy by molecular breeding.

罂粟作为鸦片及海洛因的毒品原植物，种植受到严格管控。目前，有关罂粟高特异性分子标记筛选、DNA多态性、遗传多样性、高密度连锁图谱构建以及系统进化关系的研究均罕见报道。本研究拟采用比较基因组学方法和测序技术，从系统进化、基因家族聚类、重要药用成分合成通路、果实发育通路、遗传图谱构建及高特异性分子标记挖掘等多个方面进行研究，系统挖掘罂粟基因组蕴含的遗传信息。本研究将为解析核心真双子叶植物从毛茛目中分化时间推定提供理论依据，以期揭示真双子叶植物基部类群的起源和演化、罂粟科基因组重复与近缘物种分化的关系、罂粟在毛茛目植物中的进化地位以及罂粟基因组大小的演化机制等科学问题。同时，针对不同罂粟亚种开发相应的分子标记，为后期的罂粟种属的鉴定、毒品产地来源的鉴定提供理论与实验基础。此外，通过揭示罂粟重要药用成分代谢通路及产油调控基因作用机制，为利用分子育种方法提高罂粟药用成分及油料含量提供分子依据。

罂粟是全球重要的药用植物之一，也是三大毒品原植物之一。通过全基因组测序技术对其主要生物碱代谢通路及关键基因进行研究，进一步采用分子育种来开发不产吗啡而生产具有其他重要药用成分或高产吗啡的罂粟新品种是罂粟遗传育种的重要研究方向。此外，对罂粟高特异性种间区分位点进行开发筛选，可弥补国内外法庭科学中还未有完善的罂粟种间鉴定区分方法的不足，极大提高罂粟案件物证鉴定的准确性和效率。.本研究利用二代测序技术，获得了一个2.62Gb中国本土罂粟（CHM）参考基因组，总测序量为956.96Gb，覆盖深度279.10×，contig N50为86.04kb，scaffold N50为1.25Mb，注释得到了79,668个蛋白编码基因。基于Hi-C辅助组装技术以及已发表的国外高那可丁罂粟品种（HN1）三代测序基因组数据，最终将本研究获得基因组挂载到了11条拟染色体上，最长scaffold为287.96Mb，superscaffolds N50为227.38Mb。本研究基于已发表的cDNA序列，在CHM基因组中重建了BIA代谢途径，并发现了通路上3′OHase，3′OMT及BBE基因的多个候补基因。此外，本研究通过对CHM和HN1的多部位和发育阶段的转录组比较，发现两者间存在着与农艺性状相关的基因结构差异，这些差异对罂粟新品种的形成具有重要促进作用。本研究的罂粟基因组学和转录组学分析结果将为今后罂粟分子育种研究和罂粟植物源性药品开发研究提供重要参考。另一方面，本研究开发获得了罂粟188724个SSR位点，随机选取了其中260个位点进行了特异性多重筛选，最终获取到一系列可对罂粟及其虞美人，野罂粟和鬼罂粟等近缘种进行准确区分的特异性SSR位点。为法庭科学中罂粟定性荧光标记复合扩增体系的建立提供了充足的分子标记基础。