The Hairy Roots Induction Of Echium Plantagineum And Its Transcriptome Analysis To Explore Genes Positively Regulating Alkannin/shikonin Biosynthesis

The alkannin/shikonin and their derivatives are important natural active compounds accumulated in the roots of the Lithospermeae plants.These compounds are not only famous natural pigments,but also important medicinal natural products with anti-bacterial,anti-inflammatory,and wound healing properties.Currently,studies on shikonin derivatives are mainly focused on Lithospermum erythrorhizon,Arnebia euchroma,Onosma paniculatum and other Lithospermeae plants.Echiunm plantagineum is a herbaceous shrub in the genus Echium(Borchinaceae).Previous studies have found that both the root of Echiunm plantagineum and the callus induced form Echiunm plantagineum can synthesize shikonin compounds.Accordingly,based on the existing shikonin production background of our research group,we induced the hairy roots of E.plantagineum,and analyzed the transcriptome of E.plantagineum to explore the genes possibly positively regulating alkannin/shikonin biosynthesis.The main results are as follows:1.In this study,the hairy roots of E.plantagineum was successfully induced by Agrobacterium rhizogenes ATCC15834.It provides a basis for the production in alkanin/shikonin metabolic engineering and E.plantagineum transgenic studies.2.RNA-seq was used to analyze different samples of E.plantgineum.A total of 121004 Unigenes were obtained by de novo assembly and N50 length is 2.1 kb.By Gene Ontology method,protein pathways analysis and Unigenes expression analysis,1088 Unigenes which may be involved in the synthesis of shikonin were filtered.We chose(1)G8H which may belong to alkannin/shikonin biosynthesis pathway and(2)120PR involved in the methyl jasmonate metabolic pathway for further studies to investigate their possible roles in regulating the formation of alkannin/shikonin.3.G8H-CL14070.1 and 12OPR-CL430.3 were obtained from comparison of gene expression and evolutionary analysis.The full-length CDS sequences were obtained by full length amplification.In conclusion,this study will lay a foundation for the study on biosynthesis and regulation of shikonin derivatives in E.plantagineum,and has certain reference value for the development of the shikonin metabolic engineering.