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Hibiscus mutabilis is native to China with a long cultivation history, and is an ancient garden tree species and medicinal plant. In this study, we selected three cultivars of H. mutabilis in a hybrid combination(H. mutabilis cv. Danbanbai, H. mutabilis cv. Jinqiusong, H. mutabilis cv. Mudanfen)to investigate evolutionary characteristics between the cultivars of H. mutabilis and its related species, and clarify the phylogenetic relationship between the cultivars of H. mutabilis and its related species, as well as explore the genetic model of chloroplast genome(cpDNA)of H. mutaibilis at the same time. We first sequenced the three cultivars of H. mutaibilis using Illumina NovaSeq. After assembly and annotation, three complete chloroplast genome sequences were obtained. The cpDNAs of the related species H. taiwanensis from our group, and H. syricus and H. rosa-sinensis from the gene bank. Then we carried out comparative analysis on composition and structure of cpDNAs of four species of Hibiscus and three cultivars of H. mutabilis, and completed its phylogenetic tree reconstruction. The results were as follows: (1)Total sizes of cpDNAs of H. mutabilis cv. Danbanbai, H. mutabilis cv. Jinqiusong, H. mutabilis cv. Mudanfen were 160 880, 160 879, 160 920 bp, respectively, and the total gene number was 130, including 85 protein-coding genes, eight ribosomal RNAs, and 37 transfer RNAs. (2) The comparative analyses showed that the cpDNAs of three cultivars of H. mutabilis and the related species H. taiwanensis were highly conserved, and the inverted repeat regions(IR)were all 26 300 bp; H. rosa-sinensis and H. syriacus shrank to at 25 745 and 25 598 bp, respectively.(3)The phylogenetic analysis revealed that the three cultivars were planted into a monophyletic branch, and then together with H. taiwanensis into a high support branch, indicating that H. mutabilis and H. taiwanensis had the closest relationship; Compared with H. syriacus and H. rosa-sinensis, H. mutabilis and H. taiwanensis were more closely related to H. hamabo, H. tiliaceum and H. canabinus.(4)Three cultivars of H. mutabilis could be distinguished by cpDNA sequence, the length of LSC/SSC of H. mutabilis cv. Danbanbai, H. mutabilis cv. Jinqiusong, H. mutabilis cv. Mudanfen were 89 355 bp/18 925 bp, 89 353 bp/18 926 bp, 89 400 bp/18 920 bp, respectively. And candidate molecular markers and DNA barcodes had been developed from repeat sequence and nucleotide diversity analyses, which could be used as a tool for cultivars identification.(5)The cpDNAs of H. mutabilis cv. Danbanbai and H. mutabilis cv. Jinqiusong showed a minimum difference and had the closest phylogenetic relationship. According to the relationship between their female and offspring, the maternal genetic characteristics of the cpDNAs of Hibiscus were proved. This study will help us to understand the evolutionary characteristics and phylogenetic relationship of cpDNAs of three cultivars of H. mutabilis and H. taiwanensis, and provide basic data on cpDNA for accurate identification of the cultivars of H. mutabilis and breeding of excellent cultivars. [ABSTRACT FROM AUTHOR]