Backgroud: Amaranthus palmeri, A. tuberculatus and A. arenicola are alien invasive dioecious amaranths originated from North America which have similar morphology and complex taxonomic relationship with their relatives. To search for effective molecular methods and accurate species boundary for detecting the alien invasive species, we sequenced whole chloroplast genome of 6 amaranths species, of which A. palmeri , A. arenicola , A. retroflexus and A. dubius are the first reports.Results: The complete chloroplast genome of 6 species has a circular molecular structure of 150,454 to 150,939 bp in length with 36.6% of GC content and contains a total of 134 genes, including 89 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. There are a total of 802 parsimony-informative (PI) sites within genes and intergenic spacers. The rpl22-rps19 , ndhG-I , rpl 32- trnLUAG , trnPUGG-psaJ and ccsA - ndhD are the hotspots in the genus. And the 1,601 bp fragment from rpl32 to psaC has contained maximum variants with 82 PI sites. A. arenicola differs from A. tuberculatus with 19 PI sites located in 14 genes and spacers separately. The regions for differentiate A. dubius , A. hypochondriacus and A. caudatus of the Hybrid complex only fasten on 2 coding genes and 5 intergenic spacers. The patristic distances (0.00001-0.00005) among the three species are approximate to the distance (0.00005) between individuals of A. tuberculatus . Conformed to dioecious and monoecious distinctions but different with previous phylogenetic studies, A. palmeri clustered with A. arenicola and A. tuberculatus and formed a stable clade of subgen. Acnida .Conclusion: The chloroplast genome has played a role in offering enough information for discrimination and phylogenetic relationship among the Amaranthus subgen. Acnida . The most valuable regions of chloroplast genome in Amaranthus are intergenic spacers and could differentiate A. arenicola from A. tuberculatus better. Subsequently, much more Amaranthus species should be sequenced and analyzed complementally in the future.