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Abstract Background Powdery mildew, caused by Eeysiphe heraclei, seriously threatens Heracleum moellendorffii Hance. Plant secondary metabolites are essential to many activities and are necessary for defense against biotic stress. In order to clarify the functions of these metabolites in response to the pathogen, our work concentrated on the variations in the accumulation of secondary metabolites in H. moellendorffii during E. heraclei infection. Results Following E. heraclei infection, a significant upregulation of coumarin metabolites—particularly simple coumarins and associated genes was detected by RNA-seq and UPLC-MS/MS association analysis. Identifying HmF6’H1, a Feruloyl CoA 6’-hydroxylase pivotal in the biosynthesis of the coumarin basic skeleton through ortho-hydroxylation, was a significant outcome. The cytoplasmic HmF6’H1 protein was shown to be able to catalyze the ortho-hydroxylation of p-coumaroyl-CoA and caffeoyl-CoA, resulting in the formation of umbelliferone and esculetin, respectively. Over-expression of the HmF6’H1 gene resulted in increased levels of simple coumarins, inhibiting the biosynthesis of furanocoumarins and pyranocoumarins by suppressing PT gene expression, enhancing H. moellendorffii resistance to powdery mildew. Conclusions These results established HmF6’H1 as a resistance gene aiding H. moellendorffii in combatting E. heraclei infection, offering additional evidence of feruloyl-CoA 6’-hydroxylase role in catalyzing various types of simple coumarins. Therefore, this work contributes to our understanding of the function of simple coumarins in plants’ defense against powdery mildew infection.

The powdery mildew caused by Eeysiphe heraclei is a serious concern in Heracleum moellendorffii Hance. Therefore, exploring the mechanisms underlying sugar efflux from host cells to the fungus during the plant–fungus interaction showed great significance. The study successfully cloned HmSWEET8 and HmSTP1 genes based on RNA-seq technology. The complementation assays in yeast EBY.VW4000 found HmSWEET8 and HmSTP1 transporting hexose. Over-expressing or silencing HmSWEET8 in H. moellendorffii leaves increased or decreased powdery mildew susceptibility by changing glucose concentration in infective sites. Meanwhile, over-expressing HmSTP1 in H. moellendorffii leaves also increased powdery mildew susceptibility by elevating the glucose content of infective areas. Additionally, HmSTP1 expression was up-regulated obviously in HmSWEET8 over-expressed plants and inhibited significantly in HmSWEET8 silenced plants. Co-expressing HmSWEET8 and HmSTP1 genes significantly increased powdery mildew susceptibility compared with over-expressed HmSWEET8 or HmSTP1 plants alone. The results demonstrated that HmSTP1 may assist with HmSWEET8 to promote E. heraclei infection. Consequently, the infection caused by E. heraclei resulted in the activation of HmSWEET8, leading to an increased transfer of glucose to the apoplasmic spaces at the sites of infection, then, HmSTP1 facilitated the transport of glucose into host cells, promoting powdery mildew infection.

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.