基于叶绿体基因rbcL和trnL-F序列的河北省野生苔草属亲缘关系分析

Genetic Relationship Analysis of Wild Carex Species in Hebei ProvinceBased on Sequences of Chloroplast Genes rbc L and trn L-F

为研究河北省野生苔草属(Carex)植物的亲缘关系,本研究以43种河北省野生苔草及国外引种的27份苔草种质为试验材料,克隆了叶绿体基因rbcL和trnL-F序列,进行遗传多样性及亲缘关系分析,并构建了系统发育树。结果表明,从70份苔草属植物中成功克隆到的rbcL和trnL-F序列数量分别为67条、60条,测序成功率分别为95.7%、85.7%,经正反序列拼接对齐获得序列长度分别为716 bp、976 bp,并上传至NCBI获得登录号。二者合并得到59条序列,长度为1692 bp。rbcL,trnL-F以及合并序列分别定义了43,56,58个单倍型。基于合并序列的59份种质间遗传距离为0~0.0421,平均遗传距离为0.0142,整体划分为3大类,与形态学分类基本相符。trnL-F序列有望开发为DNA条形码,以期为苔草属的分类鉴定、系统发育等研究提供依据。

To study the genetic relationships of wild Carex plants in Hebei Province,43 species of wild Carex plants in Hebei Province and 27 germplasms of Carex introduced from abroad were used as experimental materials.The chloroplast genes rbc L and trn L-F sequences were cloned for genetic diversity and phylogenetic analyses.And a phylogenetic tree was constructed.The results showed that the number of rbc L and trn L-F sequences successfully cloned from 70 plants of the genus Carex was 67 and 60,with sequencing success rates of 95.7%and 85.7%,respectively.The sequence lengths obtained after forward and backward sequence splicing and alignment were 716 bp and 976 bp,respectively and were uploaded to NCBI to obtain the GenBank accession number.The rbc L and trn L-F sequences combination obtained 59 sequences with a length of 1692 bp.The rbc L,trn L-F and combined sequence define 43,56 and 58 haplotypes.Based on the combined sequence,the genetic distance between 59 germplasms is 0~0.0421,with an average genetic distance of 0.0142.Based on NJ phylogenetic tree of combined sequence,59 germplasms could be divided into 3 groups,which was basically consistent with morphological classification.The trn L-F sequence is expected to be developed as DNA barcodes,in order to provide basis for the classification,identification and phylogenetic research of Carex.