中日产川芎的matK、ITS基因序列及其物种间的亲缘关系

matK AND ITS NUCLEOTIDE SEQUENCING OF CRUDE DRUG CHUANXIONG AND PHYLOGENETIC RELATIONSHIP BETWEEN THEIR SPECIES FROM CHINA AND JAPAN

目的 分析中国产川芎LigusticumchuanxiongHort.及日本产川芎CnidiumofficinaleMakino的核基因组ITS和叶绿体基因组matK序列 ,为探讨中日产川芎物种间的亲缘关系提供分子依据。方法 采用PCR直接测序技术测定川芎和日本川芎的ITS基因和matK基因核苷酸序列并作序列变异分析。结果 川芎和日本川芎的matK序列长度均为 12 68bp ,编码 4 2 2个氨基酸。ITS1 5 8S ITS2序列长度均为 699bp ,其中 18SrRNA基因 3′端序列 5 4bp ,ITS1序列 2 15bp ,5 8SrRNA基因序列 162bp ,ITS2序列 2 2 2bp ,2 6SrRNA基因 5′端序列 4 6bp。根据排序比较 ,川芎原植物与其商品药材间的matK基因和ITS基因序列完全相同 ,而川芎与日本川芎间matK基因则仅有 1个变异位点 ,即在上游 95 9nt处 1个转换替代 (T→C) ,反映在氨基酸序列则发生一个非同义取代V(GTG)→A(GCG) ;ITS基因也仅有 1个变异位点 ,即在ITS1上游 5 4nt处 1个转换替代 (T→C)。结论 通过进化速率较快的基因序列同源性分析 ,基本可以认为中日所产川芎基原一致 ,日本川芎学名似应改为LigusticumchuanxiongHort.

AIM To provide more molecular evidences for species relationship between Chuanxiong ( Ligusticum chuanxiong Hort.) from China and Japanese Chuanxiong (Senkyu in Japanese) ( Cnidium officinale Makino). METHODS To sequence such two genes as internal transcribed spacer (ITS) from nuclear rDNA and maturase for lysine ( mat K) in tRNA lys (UUU) intron from chloroplast DNA of both Ligusticum chuanxiong and Cnidium officinale using PCR direct sequencing and to analyze the sequence variation of two genes between these two species. RESULTS The mat K gene sequence of Ligusticum chuanxiong and Cnidium officinale is 1268 bp in length, coding 422 amino acids of maturase protein. ITS gene sequence 699 bp, consisting of 54 bp of 18S rRNA 3′, 215 bp of ITS1, 162 bp of 5 8S rRNA, 222 bp of ITS2, 46 bp of 26S rRNA 5′. Multiple sequence alignment shows that the sequence of two genes between dried crude drug and fresh voucher material of Ligusticum chuanxiong and Cnidium officinale , there is 1 variable site (T→C) in mat K (upstream at 595 nt) and ITS (ITS1 at 54 nt) between Ligusticum chuanxiong and Cnidium officinale . CONCLUSION Based on homology analysis of two genes plastid mat K and nuclear ITS, the origin of Chuanxiong from China and Japan ought to be identical, the scientific name Cnidium officinale of Japanese Chuanxiong should be changed to Ligusticum chuanxiong .