Identification and analysis of CYP450 and UGT supergene family members from the transcriptome of Aralia elata (Miq.) seem reveal candidate genes for triterpenoid saponin biosynthesis

Abstract

Background: Members of the cytochrome P450 (CYP450) and UDP-glycosyltransferase (UGT) gene superfamily have been shown to play essential roles in regulating secondary metabolite biosynthesis. However, the systematic identification of CYP450s and UGTs has not been reported in Aralia elata (Miq.) Seem, a highly valued medicinal plant. Results: In the present study, we conducted the RNA-sequencing (RNA-seq) analysis of the leaves, stems, and roots of A. elata, yielding 66,713 total unigenes. Following annotation and KEGG pathway analysis, we were able to identify 64 unigenes related to triterpenoid skeleton biosynthesis, 254 CYP450s and 122 UGTs, respectively. A total of 150 CYP450s and 92 UGTs encoding > 300 amino acid proteins were utilized for phylogenetic and tissue-specific expression analyses. This allowed us to cluster 150 CYP450s into 9 clans and 40 families, and then these CYP450 proteins were further grouped into two primary branches: A-type (53%) and non-A-type (47%). A phylogenetic analysis of 92 UGTs and other plant UGTs led to clustering into 16 groups (A-P). We further assessed the expression patterns of these CYP450 and UGT genes across A. elata tissues, with 23 CYP450 and 16 UGT members being selected for qRT-PCR validation, respectively. From these data, we identified CYP716A295 and CYP716A296 as the candidate genes most likely to be associated with oleanolic acid synthesis, while CYP72A763 and CYP72A776 were identified as being the most likely to play roles in hederagenin biosynthesis. We also selected five unigenes as the best candidates for oleanolic acid 3-O-glucosyltransferase. Finally, we assessed the subcellular localization of three CYP450 proteins within Arabidopsis protoplasts, highlighting the fact that they localize to the endoplasmic reticulum. Conclusions: This study presents a systematic analysis of the CYP450 and UGT gene family in A. elata and provides a foundation for further functional characterization of these two multigene families.

The Manchurian Walnut Genome: Insights into Juglone and Lipid Biosynthesis

“…Expression analysis showed that the obtained CYP450 genes maintain a high expression level during the differentiation of walnut exocarp, which would contribute to juglone accumulation. This result is similar to the secondary metabolite biosynthesis in plants such as Aconitum vilmorinianum (aconite) [ 60 ], Salvia miltiorrhiza (red sage) [ 61 ], Scutellaria baicalensis (Chinese skullcap) [ 62 ], and Aralia elata (angelica tree) [ 63 ]. Additionally, we identified the expanded CYP450 gene family from the genome assembly of the J. mandshurica , suggesting that they may specifically affect juglone biosynthesis.…”

Identification of Putative Candidate Genes from Galphimia spp. Encoding Enzymes of the Galphimines Triterpenoids Synthesis Pathway with Anxiolytic and Sedative Effects

“…In that work, the authors proposed a biogenetic pathway for celastrol in which hydroxyfriedelanes synthesis was mediated by P450 [ 26 ]. On the other hand, in Aralia elata , CYP716A295 and CYP716A296 were identified as the candidate genes most likely associated with oleanolic acid synthesis [ 27 ].…”

The tomato cytochrome P450 CYP712G1 catalyses the double oxidation of orobanchol en route to the rhizosphere signalling strigolactone, solanacol

“…To construct a phylogenetic relationship for the CYP712 gene family, homologous protein sequences were mined from genome assemblies of 14 species across the flowering plants (Dataset S1) and new transcriptomes from tobacco roots (Short Read Archive (SRA) accession number SRR7540357), Astragalus sinicus (SRA accession number SRR13286078) and Cosmos bipinnatus (SRA accession number SRR3546768-9) were assembled de novo using the TRINITY pipeline (Grabherr et al, 2011;Qin et al, 2020). We also included available sequences from Bupleurum chinense and Aralia elata (Araliaceae) as representatives of the Apiales order and the putative friedelin C-29 oxidase sequence from Maytenus ilicifolius (Sui et al, 2011;Bicalho et al, 2019;Cheng et al, 2020). Some genes of Kingdonia uniflora (KAF6168288), Nyssa sinensis (KAA8520263) and Trifolium pratense (Tp57577_TGAC_v2_mRNA4964) were manually re-annotated based on homology with other known Ó 2022 The Authors New Phytologist Ó 2022 New Phytologist Foundation.…”