目的 识别当归Angelica sinensis根药用活性物质的基因序列,阐释当归次生代谢产物生物合成的遗传基础.方法 从当归头、当归尾提取总RNA,质量鉴定合格后构建测序文库.测序文库检测合格后在Illumina HiSeq 2000测序仪上完成双端测序.采用相关生物信息学方法分析当归根的转录组特性.结果 采用Illumina HiSeq 2000高通量测序获得当归根转录组的66 431 540个原始短读序.用生物信息学方法从头组装并注释了30 432个功能基因序列(unigene).应用Uniprot蛋白数据库进行序列相似性比对,当归功能基因序列主要分布于目前研究较深入的7种重要经济作物,如葡萄、蓖麻、毛果杨、大豆、苜蓿、拟南芥和烟草.研究发现,当归根表达的127、69、70、94个unigene分别映射到类苯基丙烷生物合成、N-聚糖生物合成、黄酮类化合物生物合成和叶酸生物合成等路径,可能参与药用活性物质阿魏酸、当归多糖、当归总黄酮和叶酸的生物合成.结论 参与当归根重要药效物质(阿魏酸、当归多糖、当归总黄酮等)生物合成的功能基因序列可能是当归补血、活血功效的分子生物学基础.

Objective To explore the hereditary basis of secondary metabolic biosynthesis in Angelica Sinensis Radix (the roots of Angelica Sinensis) and identify the gene sequence of active substance in Angelica Sinensis Radix. Methods The samples of Angelica Sinensis Radix were collected in Min County of Gansu Province and were identified by relevant experts. Total RNA was extracted from the heads and tails of Angelica sinensis. Sequencing library was constructed and qualified. Then, Double-ended sequencing was accomplished in Illumina HiSeq 2000 device. Transcriptome characteristic of Angelica Sinensis Radix was analyzed by relevant bioinformatics. Results We obtained 66 431 540 primitive sequences of Angelica Sinensis Radix by Illumina HiSeq 2000 high-flux sequencing and annotated 30 432 unigenes by bioinformatics. The important economic crop distributed in Angelica Sinensis Radix sequences are Vitis vinifera, Ricinus communis, Populus trichocarpa, Glycine max, Medicago truncatula, Arabidopsis thaliana, and Nicotiana tabacum through comparing with 939 species sequences in Uniprot protein library. It was suggested that 127, 69, 70, and 94 unigenes expressed in Angelica Sinensis Radix were respectively mapped to the pathway of phenylpropanoid biosynthesis, N-glycan biosynthesis, flavonoid biosynthesis, and folate biosynthesis. These unigenes are related to the biosynthesis of important active substances, for example ferulic acid, polysaccharide, flavonoid, and folic acid in Angelica Sinensis Radix. Conclusion Unigenes explored in the present paper are involved in the biosynthesis of major pharmaceutical substances, suggesting the genetic basis of Angelica Sinensis Radix.