射干及类似药用植物叶绿体rbcL基因序列分析

RbcL sequence analysis of Belamcanda chinensis and related medicinal plants of Iris

目的 对射干Belamcandachinensis(L .)DC .,鸢尾IristectorumMaxim .,野鸢尾I .dichotomaPall.,蝴蝶花I.japonicaThunb .和德国鸢尾I.germaicaL .等 5种药用植物进行叶绿体rbcL基因序列分析 ,并对其亲缘关系进行探讨。方法 CTAB(Cetyltrimelhylammoniumbromide,CTAB)法提取总DNA ,用作者设计的引物对鸢尾科 5种药用植物的叶绿体rbcL(ribulose 1,5 bisphosphatecarboxyloseLargeGene ,rbcL)基因进行扩增 ,PCR扩增产物纯化后 ,用ABI3 10DNA自动测序仪测序。结果 获得射干和 4种鸢尾属药用植物叶绿体rbcL基因部分序列 (约 75 0bp) ,除德国鸢尾外 ,其余 4种药用植物的rbcL基因序列为首次获得 ;用clustal 8 0 ,MEGA 2 0等软件分析统计获得的目的基因片段 ,得到碱基突变点 ,遗传距离 [碱基差异数 ( 1 0 0 0～ 2 0 0 0 0 )、颠换数为 ( 0 0 0 0～ 9 0 0 0 )、转换数为 ( 0 0 0 0～ 14 0 0 0 ) ],根据rbcL基因部分序列数据建立分子系统树。结论 根据叶绿体rbcL基因序列数据可以很好的鉴别

Aim To identify “Shegan” [ Belamcanda chinensis (L.) DC.] and relative medicinal plants of Iris including Iris tectorum Maxim., I.dichotoma Pall., I.germanica L. and I.japonica Thunb. by ribulose 1,5 bisphosphate carboxylase Large Gene ( rbcL ) sequence analysis. Methods General DNA was isolated from the fresh leaves of Belamcanda chinensis and 4 Iris spp . by CTAB. A pair of primers was designed to amplify the rbcL gene and PCR Preps DNA kit was used to purify the PCR products. The rbcL sequences were determined by ABI (Applied Biosystems Inco.) Prism 310 Genetic Analyzer. Results A fragment of about 750 bp of rbcL gene from Belamcanda chinensis and 4 Iris spp . were amplified and sequenced. The rbcL sequences of Iris tectorum, I.dichotoma Pall. and I.japonica were reported for the first time. The rbcL sequences of 5 species of Iridaceae were aligned and analyzed using Clustal (Version 8 0) and MEGA (Version 2 0.) programs. The nucleotide number of difference is from 1 000 to 20 000. The tranversions is from 0 000 to 9 000 and the transitions is from 0 000 to 14 000. Phylogenetic tree based on rbcL partial sequence data indicated that the eleven samples of 5 species clustered separately. Conclusion The sequence variation of rbcL can be used to identify Belamcanda chinensis and 4 species of relative medicinal plants of Iris . The molecular phylogenetic tree accords with the classical taxonomy.