苦蘵抗肿瘤活性成分（Physagulin R和Physalin H）的遗传基础解析和品种选育

China has the largest population in the world, and is one of the countries with high-incidence of cancer. As one of the origins of the TCM products, separation and exploration of the key components with anti-tumor efficacy from traditional Chinese medicinal materials has become a focus in modern Chinese medicine research. The suggested project aims at the genetic dissection of the newly-identified compositions of Physagulin R and Physalin H, which have high anti-tumor activity and were firstly discovered in our lab. The multiple F1 populations and a large sample of wild germplasm of Physalis angulata are to be tested with HPLC technology to establish the high throughput phenotypic data; the whole transcriptome sequencing is to be used in comprehensive evaluation of differential gene expression analysis for individuals of F1 populations; with the whole genome sequence information of close- related species of tomato and potato as reference, genome-wide SNP markers of the above populations is to be obtained on the basis of transcriptome sequencing and restriction-site associated DNA (RAD) sequencing; both QTL linkage mapping and genome-wide association analysis (GWAS) strategies are to be applied for genetic dissection. And also, the tag SNPS of Physagulin R and Physalin H related genes will be evaluated with candidate genes association mapping, and SNP marker assisted breeding system will be established in germplasm resources selection and new variety breeding of Physalis angulata.

我国是人口大国，也是肿瘤病患高发的国家之一。传统中药材的有效活性成分分离和抗肿瘤新成分发掘已成为现代中药研究的热点。本项目旨在对最近研究发现苦蘵中具有抗肿瘤活性新成分Physagulin R和Physalin H的遗传基础进行解析，并服务于药材新品种选育。研究以农作物遗传研究思路为基础，以多个苦蘵F1分离群体和野生种质大样本群体为材料；利用高效液相分离技术获得准确的Physagulin R和Physalin H表型数据；利用转录组测序进行基因表达差异分析；参考同科近缘种番茄和土豆全基因信息，运用转录组和简化基因组测序获得各群体全基因组SNP标记；据此对上述成分含量的遗传基础进行连锁定位和全基因组关联分析。再通过候选基因关联分析，深入挖掘两种方法共同解析的Physagulin R和Physalin H含量相关基因内标签SNP位点，构建分子标记辅助育种体系，用于苦蘵种质资源筛选和新品种选育。

苦蘵是常见的酸浆属植物，本项目组前期的研究发现该植物宿萼中含有的化合物Physagulin R和Physalin H具有很好的抗肿瘤活性作用。为深入了解这两种化合物的遗传基础，本项目在广泛收集酸浆属植物种质资源700余份（其中苦蘵400余份）的基础上，开发酸浆属植物的分子鉴定技术，保证了原材料的准确性；利用全基因组SNP标记分析了国内主要种质资源的遗传多样性和遗传结构特点；开发了利用分子标记进行苦蘵杂交后代真实性鉴定的体系；利用全基因组关联分析的技术，分别获得Physagulin R含量相关QTL27个，Physalin H含量相关QTL67个，为系统解析化合物遗传基础提供了候选区段；同时利用Physagulin R和Physalin H含量差异显著的种质资源开展毛状根诱导和MeJA处理的研究，从代谢组、转录组和蛋白组解析Physagulin R和Physalin H遗传代谢相关的路径和关键步骤；结合Physagulin R和Physalin H化学结构的特点，预测了其代谢通路的主要步骤及其关键酶；此外，本项目还构建了苦蘵功能基因研究的遗传转化体系，目前的效率是1.11%。本项目从抗肿瘤活性化合物出发，从化学结构、代谢通路、关键步骤的酶和影响因子多角度开展代谢组、转录组和蛋白组多组学的研究，为苦蘵乃至相关药用植物主要活性成分的研究开辟了新的思路，相关研究成果无论对基础研究还是中药产业化都具有重要的作用。