目的 以药用植物单叶铁线莲Clematis henryi叶片为材料，在高通量测序、组装和序列分析的基础上，明确叶绿体因组的序列特征和系统发育关系。方法 利用CTAB法提取叶片基因组DNA，通过NovaSeq 6000平台测序，再用NovoPlasty组装叶绿体基因组，借助PhyML生成系统发育树。结果 单叶铁线莲的叶绿体基因组全长159 707 bp，GC值为38.0%，大单拷贝区、小单拷贝区和反向重复序列的长度分别为79 449、18 100、31 079 bp；单叶铁线莲的叶绿体基因组总共有基因137个，其中蛋白质编码基因、tRNA、rRNA的数量分别为91、36、8，另有2个假基因，分别为Ψycf1和ΨinfA。序列比对结果表明，单叶铁线莲与牯牛铁线莲叶绿体基因组的相似性最高，达99.6%；共检测到41个SSR，其中40个为单核苷酸重复，1个为三核苷酸重复。系统发育分析结果表明，单叶铁线莲与牯牛铁线莲和大叶铁线莲聚于一组，支持率达100%。结论 单叶铁线莲叶绿体基因组的组装、序列分析和系统发育分析，为该药用植物的遗传结构和遗传多样性研究奠定了基础。

Objective To confirm chloroplast genome sequence characteristics and phylogenetic relationships of Clematis henryi based on high-throughput sequencing, genome assembly, and sequence analysis. Methods The CTAB-based DNA extraction method was used to isolate leaf genomic DNA, and high-throughput sequencing was performed with the HiSeq X Ten Platform. The chloroplast genome was assembled using NovoPlasty and a phylogenetic tree was generated by PhyML. Results The full chloroplast genome of C. henryi was 159 707 bp in length and its GC content was 38.0%. The sizes of large single copy, small single copy, and inverted region were 79 449 bp, 18 100 bp, and 31 079 bp, respectively. Totally, there were 137 genes, and the numbers of protein-coding genes, tRNAs, and rRNAs were 91, 36, and eight, respectively. Additionally, Ψ ycf1 and Ψ infA were recognized as two pseudogenes. Sequence comparison results showed that the highest similarity existed between C. henryi and C. guniuensis, which reached 99.6%. A total of 41 SSRs were detected, in which 40 were mono-nucleotide and one was tri-nucleotide. Phylogenetic analysis indicated that C. henryi, C. guniuensis and C. heracleifolia gathered in the same clade, with a support rate of 100%. Conclusion The assembly, sequence analysis, and phylogenetic analysis of C. henryi chloroplast genome provided insight into studies on both genetic structure and genetic diversity of this medicinal plant.

R286.12

台州市211人才工程经费资助（2012年度）