摘要小酸浆（Physalis minima），是一年生茄科（Solanaceae）酸浆属（Physalis）草本植物，具有很高的药用价值。目前，对于小酸浆种质资源的多样性仍缺乏科学规范的研究体系，致使小酸浆种间尚存在混杂。为完善小酸浆种质资源的遗传多样性数据库，本实验通过 SSR分子标记技术对来自唐山、丽水、东明、菏泽、六安、平顶山这6个中国不同地区的82个小酸浆品种进行品种遗传多样性的研究。主要研究方法和结果如下:73041

1、82个小酸浆样品基因组DNA的提取：取小酸浆鲜叶为原料，利用EZUP柱式植物基因组DNA抽提试剂盒提取小酸浆基因组DNA，并且通过1。5%的琼脂糖凝胶电泳检测其纯度，检测结果证明提取的DNA可用于进行后续实验。

2、小酸浆DNA的SSR-PCR反应体系(10μL)：DNA模板10ng，1μl，5×MIX5μl，正反向引物各0。25mol/L，0。5μl。SSR-PCR扩增程序：94 ℃，5 min预变性；<94 ℃，40 s变性，根据不同引物相应的Tm退火值（一般为56℃）40 s复性，72 ℃，1 min延伸>统共35个循环；最终72 ℃延伸10 min，4 ℃恒温保存。

3、基于SSR标记对小酸浆进同源分析：利用SSR-PCR的方法筛选出8对SSR标记对来自中国6个地区的82个小酸浆DNA进行同源性分析，总共扩出61条SSR条带，其中多态性条带有49条（占80。33%）。平均每个引物产生6。13条多态性条带。证明了SSR引物具有良好的多态性，适合于小酸浆的遗传多样性研究。

4、基于SSR标记数据的UPGMA聚类结果显示，小酸浆种质间的遗传相似系数为0。559~0。989，另外通过主坐标分析和UPGMA聚类发现， SSR标记将82份小酸浆种质资源聚类为I、II两大类，I类包括21份来自唐山的种质，II类又分为3个亚类，分别是II-1丽水、东明的28个种质，II-2菏泽的11个种质以及II-3六安、平顶山的21个种质。

毕业论文关键词：SSR标记； 小酸浆； PCR；遗传多样性；聚类分析；主坐标分析（PcoA）

Abstract Physalis minima a species of Solanaceae annual herb, has a very high medicinal value。 Currently,the study of Genetic persity is still lack of Scientific and standardized Research system , which results in Physalis minima’s Interspecific hybrid 。 In order to improve the genetic persity database of germplasm resources ,the genetic persity of 82 samples from 6 different areas (Tangshan,Lishui,Dongming,Heze,Liuan, Pingdinshan) was studied based on SSR markers 。 The main contents and results of this experiment are as follows :

1、82 Physalis minima samples’ genome DNA : The genomic DNA was isolated using the Modified Ezup pillar Plant Genomic DNA Extraction kit with fresh, young Physalis minima leaves。

2、The SSR-PCR reaction system of Physalis minima genomic DNA(10μL)：DNA template 10ng，1μl，5×MIX5μl，the forward /reverse primer 0。25mol/L，0。5μl。 SSR-PCR Amplification program 94ºC for 5 min, followed by 35 cycles of 94ºC for 40s, 50ºC (Primer Tm depends) for 40s, 72ºC for 1 min, and a final extension at 72ºC for 10 min。

3、The homology analysis of Physalis minima based on SSR mark: Use 8 pairs of SSR mark to homology analysis 82 samples from 6 different areas of China。 produced 61 reliable loci among 82 populations of Physalis minima , there are about 49 polymorphism bands (accounts 80。33%), each primer produced 6。13 reliable loci。 This result proved the SSR molecular markers are good in polymorphism , and can be used to studying Physalis minima genetic polymorphisms。

4、UPGMA cluster analysis based on SSR molecular markers data, showed that the range of genetic similarities was 0。559~0。989。 The partition of clusters based on SSR molecular markers clustered 82 Physalis minima germplasms into 2 main groups in the UPGMA dendrogram and PCoA plot。 Group I have 22 germplasms from TS。 Cluster II comprised 3 subgenera , they are II-1 include 28 germplasms from LS and DM,II-2 include 11 germplasms of HZ ,II-3 include 21 germplasms from LA and PDS。