EuRBG10 involved in indole alkaloids biosynthesis in Eucommia ulmoides induced by drought and salt stresses.

Alkaloids are natural products with many important medicinal activities. To explore the mechanism of abiotic stress promoting alkaloid biosynthesis in Eucommia ulmoides, transcriptomic analysis and metabonomic analysis were used, virus-induced gene silencing (VIGS) lines of target gene were constructed. The results showed that drought and salt stress caused wilting and blackening of leaves, decreased chlorophyll level, and significantly induced MDA and relative conductivity. To resist the damage of stress to cells, the level of secondary metabolites such as alkaloids increased significantly with the extension of stress time. Transcriptomic results showed that, were. Six alkaloid related genes (AWGs) were gathered in five modules positively correlated with either salt stress or alkaloid contents by WGCNA. Results of GO and KEGG enrichment revealed that biosynthesis of alkaloid, especially indole alkaloid was induced, and degradation of alkaloid was inhibited under salt stress. Combining the results of transcriptome and metabolomics, it was suggested that EuRBG10 promotes the production of indole alkaloids and EuAMO5 inhibits the degradation of alkaloids, which may be the core mechanism of the indole alkaloid biosynthesis pathway (map00901) induced by salt stress. The results of these hub proteins were also consistent with the chordal graph of KEGG enrichment. Hub roles of EuRGB10 was checked in E. ulmoides by VIGS. Our findings provide a preliminary understanding of abiotic stress regulating secondary metabolites such as alkaloids, and propose hub genes that can be used to improve the level of bioactive components in medicinal plant.

A beta-glucosidase gene from Stevia rebaudiana may be involved in the steviol glycosides catabolic pathway.

We herein report the preparation of a full-length raucaffricine-O-beta-D-glucosidase gene of stevia rebaudiana Bertoni (named SrRG1, GenBank accession number MK920450). Sequence analysis indicated SrRG1 consists of a 1650 bp open reading frame encoding a protein of 549 amino acids. Its deduced amino acid sequence showed a high identity of 82% with a raucaffricine-O-beta-D-glucosidase from H. annuus of glycoside hydrolase family 1. The expression pattern analyzed by real-time quantitative PCR showed no significant difference among different tissues, developmental stages, and cultivars under normal growth conditions. Furthermore, the gene function of SrRG1 was preliminarily studied by agrobacterium-mediated transformation on instantaneous expression. In the test of agrobacterium-mediated transformation on instantaneous expression, it was observed that overexpression of SrRG1 increased the accumulation of steviol content and decreased the major components and total SGs contents. Such results demonstrated that SrRG1 may participate in the steviol glycosides catabolic pathway. However, the effect of silencing construct infiltration on steviol and SGs content was not significant and its expression pattern was constitutive, which most probably, attributed the hydrolysis of SGs to the secondary activity of SrRG1. This study firstly identified the bate-glucosidase in stevia and advances our understanding of steviol glycosides hydrolyzation.