Distinguish the tribe Podophylleae species using molecular marker based on chloroplast genomes

Podophyllotoxin (PPT) is widely used to semi-synthesize derivatives for chemotherapy agents against cancers. Presently, PPT mainly exists in tribe Podophylleae that are composed of 4 genera with 13 species. To rationally develop the tribe Podophylleae, it is necessary to distinguish the 13 species and avoid the confusion in medicinal market, while a rapid and efficient approach is limited based on molecular markers. Here, chloroplast genome of new Dysosma villosa species was sequenced, differences in chloroplast genomes among the 13 tribe Podophylleae species were analyzed, and then molecular makers for distinguishing the tribe Podophylleae species were screened. The results showed that the new D. villosa species contained 157,358 bp with encoding 128 genes and had a close relationship with D. delavayi . There were 4 unique genes ( e.g. , psbL , rps19 , and infA ) and 13 highly variable genes [ e.g. , ndhK (NADH dehydrogenase subunit K), trnA- UGC (tRNA-Alanine with anticodon UGC), and trnD -GUC] in the chloroplast genomes among the 13 tribe Podophylleae species. Eight molecular markers ( e.g. , trnR -UCC, trnA -UGC, and ndhK ) were successfully validated as species-specific tools to distinguish 6 target species ( e.g. , S. hexandrum , D. aurantiocaulis , and D. villosa ) with the method of agarose gel electrophoresis or base sequencing. These findings complement previous studies on molecular markers to identify the tribe Podophylleae and will provide useful references to enhance artificial cultivation of high-yield PPT species. • Chloroplast genome of D. villosa contains 157,358 bp with 18,976 codons and 68 SSRs. • D. villosa has a close genetic relationship with D. delavayi based on phylogenetic analysis. • Four unique genes ( e.g. , psbL , rps19 , and infA ) were observed in the 13 tribe Podophylleae. • Thirteen highly variable genes ( e.g. , ndhK , trnA -UGC, trnD -GUC) were observed. • Eight genes are validated as molecular markers to distinguish 6 tribe Podophylleae species.