[Objective] The research aimed to distinguish Chinese herb Qingjiao from its botanical origin plants by comparing different DNA sequences,so as to provide a molecular basis for origin identification and quality evaluation.[Method] The cpDNA psbA-trnH and nrDNA ITS sequences of five Chinese herb Qingjiao plants,including Gentiana macrophylla pall.,Gentiana straminea Maxim.,Gentiana crassicaulis Duthie ex Burk.,Gentiana dahurica Fisch and Gentiana officinalis H.Smith,were amplified with PCR,and then sequenced by direct PCR sequencing method for homologous analysis.[Results] The length of cpDNA psbA-trnH of five plants was 316-318 bp;there were seven different haplotypes and seven variable sites;the GC content of the sequence was 21.2%;the phylogenetic clustering showed the same result as haplotype analysis.The length of nrDNA ITS sequence of five plants was 624-625 bp,there were five different haplotypes and 13 variable sites;the GC content of the sequence was 59.3%.The result of phylogenetic clustering suggested that G.dahurica and G.straminea,G.macrophylla and G.officinalis clustered together as sister clades,respectively.[Conclusion] The nucleotide differences of nrDNA ITS regions could be used for distinguishing botanical origin in Chinese herb Qingjiao.