【摘要】 基于高通量的转录组测序,为大量并快速的开发分子标记提供契机。本研究对紫草科植物疏花软紫草进行转录组测序,共得到92 042 086条reads,从疏花软紫草158 446个Unigene中搜索到332个低拷贝核基因。为进行多态性验证,从中随机挑选30个设计引物,并对3个疏花软紫草种群共9个个体进行PCR扩增,结果发现其中16对引物能够扩增出单一稳定的目的条带,从中随机挑选7个片段进行群体测序并计算DNA多态性。结果表明,7对引物的多态性位点平均值为6.57,单倍型平均值为5.14,单倍型多态性(HD)在0.389~0.944之间,核苷酸多态性(Pi)为0.001 48~0.017 23。通过转录组测序技术开发的低拷贝核基因有较高的多态性,且可以有效地运用于群体遗传学和谱系地理学研究中。另外,这些低拷贝核基因能够被运用于软紫草属植物系统发育重建以及物种形成等研究工作中。更多还原

【Abstract】 Transcriptome sequencing based on high throughput has provided an opportunity for rapidly developing more molecular markers. 92 042 086 reads were obtained in this research based on transcriptome sequence of Arnebia szechenyi,then acquiring 332 low-copy genes by searching 158 446 unigene of A. szechenyi. In order to verify DNA polymorphism,this research randomly selected 30 low-copy genes as designing primers and 9 individuals in three natural populations of A. szechenyi were detected by PCR amplification. It was found that 16 pairs of 30 low-copy gene primers could amplify steadily single target band. After that,7 fragments randomly selected from 16 primers were sequenced and used to calculate DNA polymorphism,the results showed that mean value of polymorphic sites based on 7 primers is 6. 57,hyplotype is 5. 14,hyplotype diversity( HD) was from 0. 389 to 0. 944,nucleotide diversity( Pi) was from 0. 001 48 to 0. 017 23.Our findings indicated that low-copy nuclear genes developed in this study based on transcriptome sequencing have relatively high polymorphism,and it could be validly used for the study of populations genetic and phylogeography. Furthermore,these low-copy nuclear genes could be applied to the fields of phylogeny reconstruction and speciation of Arnebia.更多还原