中国不同地区蛇床的rDNA ITS序列分析

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

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Ｐ５－８ＳＩＴＳ１＋ Ｐ５－８ＳＩＴＳ２ ，ＰＣＲ扩增产物纯化后用银染法或ＡＢＩ３１０ 测序。结果：得到核糖体ＤＮＡ中的ＩＴＳ及５－８ＳｒＤＮＡ 完全序列，１８Ｓ和２６ＳｒＤＮＡ 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。

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 .