Pharmacological Research - Modern Chinese Medicine (Mar 2022)
De nove transcriptome sequencing identifies genes involved in aconitine-type alkaloids biosynthesis in Aconitum kusnezoffii Reichb.
Abstract
Ethnopharmacological relevance: Aconitum kusnezoffii Reichb. is one of the plants of Aconitum genus, and it′s root, flower, bud, and leaf has been used for traditional Mongolian medicine to bactericidal, relieve pain, collapse, rheumatic fever and so on, according to the Pharmacopoeia of the People's Republic of China (2021 edition). Aim of the study: We aimed to reveal genes involved in the biosynthesis and accumulation of the three major medicinal components (aconitine, mesaconitine and hypaconitine) of Aconitum kusnezoffii Reichb. Materials and Methods: Aconitum kusnezoffii Reichb. samples of root, flower, leaf, and stem were collected for de novo transcriptome sequencing and bioinformatics analysis. Dry samples of root, flower, leaf, and stem were used for quantitative analysis of aconitine, mesaconitine and hypaconitine by HPLC analysis. Results: the HPLC results show the content of aconitine, mesaconitine and hypaconitine was high in the root, then in flower, and low in leaf and stem. Mesaconitine was the most abound in all the four tissues. Transcriptome sequencing analysis shows a total of 145,117 unigenes were obtained, and 38,540 of the unigenes have complete CDS. Differential analysis indicated secondary metabolite related pathways were significantly enriched in flower, including diterpenoid, alkaloid, and flavonoid biosynthesis pathway. And 55 genes related to aconitine, mesaconitine and hypaconitine biosynthesis were identified, the expression analysis showed most of the genes were low expressed root, but high expressed in flowers. Nearly 2000 genes were correlated expressed to 5 high expressed GGPS and 1 CPS genes, and 12 genes had protein-protein interaction involved in aconitine, mesaconitine and hypaconitine biosynthesis. Conclusion: We performed de novo transcriptome analysis of four different tissues of Aconitum kusnezoffii Reichb., and identified the genes involved in aconitine, mesaconitine and hypaconitine biosynthesis. The content of aconitine, mesaconitine and hypaconitine in the four tissues of Aconitum kusnezoffii Reichb. was measured. The aconitine, mesaconitine and hypaconitine biosynthesis genes expression and their content in plant tissues were inconsistent, indicated there exist a source-sink relation.
