中国不同地区蛇床的rDNA ITS序列分析

目的：探讨不同分布区的蛇床Cnidium monnieri的ITS序列变异与其地理分布和化学成分的相关性。方法：设计2对引物，Pf+Pb及P5.8S ITS1+P5.8S ITS2，PCR扩增产物纯化后用银染法或ABI 310测序。结果：得到核糖体DNA中的ITS及5.8S rDNA完全序列，18S和26S rDNA部分序列,共约700 bp。5个地点样品的ITS-1及ITS-2的序列大小分别为210～217 bp和219～224 bp。ITS-1碱基序列的遗传距离0.00～1.93%，ITS-2碱基序列的遗传距离0.46～2.34%，ITS-1较为保守。以NJ法根据ITS-2序列数据重建系统发生树。哈尔滨样品聚为一组，衡水与德州样品和郑州与高淳样品各自聚为一组。结论：ITS-2序列的变异与中国产蛇床的纬度分布相关，而其与蛇床化学型的关系尚需作进一步研究。

RIBOSOMAL DNA ITS SEQUENCE ANALYSES OF CNIDIUM MONNIERI FROM DIFFERENT GEOGRAPHICAL ORIGIN IN CHINA

AIM: To study the correlation between ITS sequence variation of Cnidium monnieri and geographical distribution and chemo type of the plant. METHODS: Two pairs of primers Pf Pb and P5 8S ITS1 P5 8S ITS2 were designed. The PCR products were purified and sequenced by silver staining DNA sequencing technique and ABI 310. RESULTS: Complete sequcnce of ITS and 5 8S rDNA, and partial sequence of 18S rDNA and 26S rDNA of Cnidium monnieri were obtained. They totaled about 700 bp. The sequences of ITS 1 and ITS 2 ranged from 210 bp to 217 bp and 219 bp to 224 bp; respectively. Pairwese sequence divergences of the samples ranged from 0% to 1 93% in ITS 1, and from 0 46% to 2 34% in ITS 2. The ITS 1 is more conservative. Phylogenetic tree based on ITS 2 sequence data (225 aligned sites) indicated that the Harbin samples, Hengshui and Dezhou samples, and Zhengzhou and Gaochun samples clustered separately. CONCLUSION: The sequence variation of ITS 2 is correlated to latitude distribution of Cnidium monnieri in China. Further studies are needed to clarify the relation between ITS sequence variation and chemo type of Cnidium monnieri .