Ginsenosides and Biotic Stress Responses of Ginseng.

Ginsenosides are saponins that possess a sugar moiety attached to a hydrophobic aglycone triterpenoid. They have been widely studied for their various medicinal benefits, such as their neuroprotective and anti-cancer activities, but their role in the biology of ginseng plants has been much less widely documented. In the wild, ginsengs are slow-growing perennials with roots that can survive for approximately 30 years; thus, they need to defend themselves against many potential biotic stresses over many decades. Biotic stresses would be a major natural selection pressure and may at least partially explain why ginseng roots expend considerable resources in order to accumulate relatively large amounts of ginsenosides. Ginsenosides may provide ginseng with antimicrobial activity against pathogens, antifeedant activity against insects and other herbivores, and allelopathic activity against other plants. In addition, the interaction of ginseng with pathogenic and non-pathogenic microorganisms and their elicitors may trigger increases in different root ginsenosides and associated gene expression, although some pathogens may be able to suppress this behavior. While not covered in this review, ginsenosides also have roles in ginseng development and abiotic stress tolerance. This review shows that there is considerable evidence supporting ginsenosides as important elements of ginseng's defense against a variety of biotic stresses.

Development and applications of a CRISPR activation system for facile genetic overexpression in Candida albicans.

For the fungal pathogen Candida albicans, genetic overexpression readily occurs via a diversity of genomic alterations, such as aneuploidy and gain-of-function mutations, with important consequences for host adaptation, virulence, and evolution of antifungal drug resistance. Given the important role of overexpression on C. albicans biology, it is critical to develop and harness tools that enable the analysis of genes expressed at high levels in the fungal cell. Here, we describe the development, optimization, and application of a novel, single-plasmid-based CRISPR activation (CRISPRa) platform for targeted genetic overexpression in C. albicans, which employs a guide RNA to target an activator complex to the promoter region of a gene of interest, thus driving transcriptional expression of that gene. Using this system, we demonstrate the ability of CRISPRa to drive high levels of gene expression in C. albicans, and we assess optimal guide RNA targeting for robust and constitutive overexpression. We further demonstrate the specificity of the system via RNA sequencing. We highlight the application of CRISPR activation to overexpress genes involved in pathogenesis and drug susceptibility, and contribute toward the identification of novel phenotypes. Consequently, this tool will facilitate a broad range of applications for the study of C. albicans genetic overexpression.