Plant secondary metabolic pathways are finely regulated by the activity of transcription factors, among which members of the bHLH and MYB subfamilies play a main role. Cannabis sativa L. is a unique officinal plant species with over 600 synthesized phytochemicals having diverse scale-up industrial and pharmaceutical usage. Despite comprehensive knowledge of cannabinoids’ metabolic pathways, very little is known about their regulation, while the literature on flavonoids’ metabolic pathways is still scarce. In this study, we provide the first genome-wide analysis of bHLH and MYB families in C. sativa reference cultivar CBDRx and identification of candidate coding sequences for these transcription factors. Cannabis sativa bHLHs and MYBs were then classified into functional subfamilies through comparative phylogenetic analysis with A. thaliana transcription factors. Analyses of gene structure and motif distribution confirmed that CsbHLHs and CsMYBs belonging to the same evolutionary clade share common features at both gene and amino acidic level. Candidate regulatory genes for key metabolic pathways leading to flavonoid and cannabinoid synthesis in Cannabis were also retrieved. Furthermore, a candidate gene approach was used to identify structural enzyme-coding genes for flavonoid and cannabinoid synthesis. Taken as a whole, this work represents a valuable resource of candidate genes for further investigation of the C. sativa cannabinoid and flavonoid metabolic pathways for genomic studies and breeding programs.

Aim: To evaluate a label-free and variant-independent assay called direct virus capture (DVC) for detection of intact SARS-CoV-2 virions through light scattering, utilizing an optical inverted lase... Plain language summary SARS-CoV-2, the virus responsible for the COVID-19 pandemic, can spread quickly from person to person and can make people very ill. An important part of controlling the sprea...

Background/Objectives: Colorectal cancer (CRC) is one of the most aggressive malignancies and has a very high mortality rate. Several studies have shown that obesity and hyperlipidemia are among the factors implicated in the onset of this disease. These factors can be modified through lifestyle changes, and diet plays a crucial role in this context. We evaluated the effects of Tuber borchii (T. borchii) fungal extracts based on experimental evidence showing that some truffles produce antioxidant, anti-inflammatory, and anticancer secondary metabolites. Methods: To this end, we treated human colorectal cancer cells (HCT 116) with various extracts of T. borchii at different time points and concentrations. Results and Conclusions: The results showed that the treatments caused a decrease in cell proliferation due to the induction of apoptotic cell death, as evidenced by FACS analyses. The apoptotic pathway was confirmed by the increase in the cleavage of Caspase 3 and Caspase 9. We then investigated the molecular mechanisms underlying cell death, finding increased nuclear localization of p53 and increased expression of its downstream pro-apoptotic genes, PUMA and NOXA. Among the upstream signaling events, we identify an increase in p-ERK1/2, a MAPK member involved in several antiproliferative/pro-apoptotic insults.

The rhizosphere is a narrow and dynamic soil zone where bacteria and fungi establish symbiotic and synergistic relationships with each other and with plants, collectively influencing nutrient cycling, soil health, and overall ecosystem functioning. Mycorrhiza Helper Bacteria (MHB) are a diverse group of bacterial taxa that promote mycorrhiza establishment, together with those that positively influence already established symbiotic associations. MHB have been largely described from the perspectives of interaction mechanisms, ecological functions, or agricultural applications, treating them as generalized fungal growth-promoters or biofertilizers. However, how their effects change throughout fungal development has received little attention. This review addresses this knowledge gap by reframing MHB not as generic growth promoters, but as stage-specific regulators of the fungal development program. We integrate current evidence on transcriptional responses, metabolite-mediated signaling, and emerging small RNA regulatory networks across five sequential phases of the fungal life cycle: spore germination, presymbiotic hyphal growth, mycelial expansion, root colonization, and nutrient uptake. Elucidating these stage-specific regulatory mechanisms may provide the basis for the rational, stage-targeted design of MHB-based innovative strategies for sustainable agriculture.

Tuber genus includes hypogeous ectomycorrhizal (ECM) fungi, widely recognized as truffles. Truffles spend much of their life in a vegetative form in the soil, a crucial phase to ensure the growth and the accomplishment of their life cycle. Adverse events to which they may be subjected, i.e., rise in temperature, drought, and biotic stress factors, can influence the levels of oxidative stress, increasing the production of free radicals. It is known that both prokaryotes and eukaryotes have evolved various primary antioxidant defenses to protect themselves from oxidative damage. In some eukaryotes, transcription factors are activated to regulate the genes responsible for producing enzymes and proteins necessary to reduce ROS levels. In yeasts the Yap1 protein is a key modulator of oxidative stress response. In this study, we used an in silico approach to characterize the Yap1 transcription factor in fungi of the Tuber genus, using Saccharomyces cerevisiae as the main reference model. The phylogenetic analysis based on the Yap1 protein sequences from 24 fungal species showed clustering consistent with their taxonomic relationships, including those observed within Tuber genus. The amino acid alignment revealed the conservation within the Tuber species of cysteine-rich domains (nCRD and cCRD) and motifs potentially involved in redox regulation. Finally, by silico investigation of Yap1 possible protein interactions and three-dimensional structure in Tuber melanosporum, it was possible to provide further elements to understand the molecular mechanisms underlying the functionality of Yap1 and the response mechanism to oxidative stress modulated by it in fungi of the Tuber genus. The evidence obtained represents an essential starting point for understanding a complex process that can strongly influence the biological cycle of the truffle and compromise its fruiting.