Abstract
BACKGROUND: Aconitum carmichaelii (AC), a traditional Chinese medicinal herb, provides substantial economic benefits to the pharmaceutical industry and rural development. Its broad genetic diversity has led to multiple cultivars with distinct biological traits. However, accurately identifying these cultivars is challenging due to the morphological similarities, particularly at the seedling stage. Misidentifying cultivars or selecting inappropriate cultivation regions may lead to crop failure. Moreover, the chemical profiles of different cultivars remain insufficiently characterized. Therefore, there is an urgent need to develop rapid and accurate authentication methods and to clarify quality-related differences among AC cultivars. RESULTS: We sequenced and analyzed the chloroplast (cp) genomes of 26 AC samples and measured the content of their major active compounds. The result revealed that the cp genomes were highly conserved, ranging from 155,880 to 155,891 bp, and contained 42–47 simple sequence repeats (SSRs), primarily mononucleotide repeats. Two DNA barcodes, trnT(GGU)-psbD 1F/1R and trnS(UGA)-psbZ 1F/1R, were identified as effective tools for differentiating between AC cultivars. Phylogenetic analyses clustered the 26 samples into three groups, Suggesting the presence of three distinct cultivars relevant to agricultural production. The divergence time of AC was estimated to be approximately 1.38 million years ago (Mya). Additionally, the total content of monoester alkaloids (benzoylaconitine, benzoylmesaconine, and benzoylhypacoitine) ranged from 0.07 to 0.12 mg/g, while diester alkaloids (aconitine, meaconitine, and hypoaconitine) ranged from 1.22 to 1.82 mg/g. Notably, the YFIII cultivars exhibited the highest levels of monoester alkaloids and lowest levels of diester alkaloids, indicating a potential safety advantage. Furthermore, 11 typical chromatographic peaks were identified through peak alignment and multi-point correction. Among them, the VIP values for P6, P8, and P9 were more significant than 1, suggesting that these peaks may serve as potential markers for quality control in AC cultivars. CONCLUSIONS: Our findings highlight AC cultivars’ significant genetic and chemical diversity. The chloroplast markers trnT(GGU)-psbD 1F/1R and trnS(UGA)-psbZ 1F/1R are practical tools for the precise identification and selective breeding of AC. This genetic diversity likely contributes to variations in the plant’s chemical composition. This study lays an important foundation for the sustainable utilization and conservation of AC genetic resources. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12870-025-07289-w.